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1. NLRX1 inhibits the early stages of CNS inflammation and prevents the onset of spontaneous autoimmunity.

Author

Marjan Gharagozloo, Shaimaa Mahmoud, Camille Simard, Kenzo Yamamoto, Diwakar Bobbala, Subburaj Ilangumaran, Matthew D Smith, Albert Lamontagne, Samir Jarjoura, Jean-Bernard Denault, Véronique Blais, Louis Gendron, Carles Vilariño-Güell, A Dessa Sadovnick, Jenny P Ting, Peter A Calabresi, Abdelaziz Amrani, and Denis Gris

Subjects
Biology (General), QH301-705.5
Abstract

Nucleotide-binding, leucine-rich repeat containing X1 (NLRX1) is a mitochondria-located innate immune sensor that inhibits major pro-inflammatory pathways such as type I interferon and nuclear factor-κB signaling. We generated a novel, spontaneous, and rapidly progressing mouse model of multiple sclerosis (MS) by crossing myelin-specific T-cell receptor (TCR) transgenic mice with Nlrx1-/- mice. About half of the resulting progeny developed spontaneous experimental autoimmune encephalomyelitis (spEAE), which was associated with severe demyelination and inflammation in the central nervous system (CNS). Using lymphocyte-deficient mice and a series of adoptive transfer experiments, we demonstrate that genetic susceptibility to EAE lies within the innate immune compartment. We show that NLRX1 inhibits the subclinical stages of microglial activation and prevents the generation of neurotoxic astrocytes that induce neuronal and oligodendrocyte death in vitro. Moreover, we discovered several mutations within NLRX1 that run in MS-affected families. In summary, our findings highlight the importance of NLRX1 in controlling the early stages of CNS inflammation and preventing the onset of spontaneous autoimmunity.

Published
2019
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2. Apelin Stimulation of the Perivascular MuSC Niche Enhances Endogenous Repair in Muscular Dystrophy

Author

Emmeran Le Moal, Yuguo Liu, Jasmin Collerette-Tremblay, Simon Dumontier, Joël Boutin, Junio Dort, Zakaria Orfi, Joris Michaud, Hugo Giguère, Alexandre Desroches, Kien Trân, François Vézina, Sonia Bedard, Catherine Raynaud, Frederic Balg, Philippe Sarret, Michelle S. Scott, Jerome N. Feige, Jean-Bernard Denault, Nicolas A. Dumont, Eric Marsault, Mannix Auger-Messier, and C. Florian Bentzinger

Abstract

Impaired skeletal muscle stem cell (MuSC) function has long been suspected to contribute to the pathogenesis of muscular dystrophy (MD). Here we describe that defects in the endothelial cell (EC) compartment of the perivascular stem cell niche in three different types of MD are associated with inefficient mobilization of MuSCs following tissue damage. Using chemoinformatic analysis, we identified the 13 amino acid form of the peptidic hormone apelin (AP-13) as a candidate for systemic stimulation of skeletal muscle ECs. In dystrophic mice, administration of AP-13 generates a pro-myogenic EC-rich niche that supports MuSC function and markedly improves tissue regeneration, muscle strength, and physical performance. Moreover, we demonstrate that EC specific knockout of the AP-13 receptor leads to regenerative defects that phenocopy major pathological features of MD. Altogether, we provide in vivo proof-of-concept that enhancing endogenous repair by targeting the perivascular niche is a viable therapeutic avenue for MD and characterize AP-13 as a novel drug candidate for systemic treatment of stem cell dysfunction.

Published
2022

3. Characterization of caspase-7 interaction with RNA

Author

Jean-Bernard Denault and Alexandre Desroches

Subjects
Proteolysis, Immunoblotting, Poly (ADP-Ribose) Polymerase-1, Apoptosis, RNA-binding protein, Cleavage (embryo), Biochemistry, Caspase 7, Substrate Specificity, 03 medical and health sciences, medicine, Humans, Molecular Biology, Caspase, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, medicine.diagnostic_test, biology, Caspase 3, Chemistry, Lysine, 030302 biochemistry & molecular biology, RNA-Binding Proteins, RNA, Cell Biology, HCT116 Cells, Recombinant Proteins, Amino acid, Cell biology, HEK293 Cells, biology.protein, Protein Multimerization, Protein Binding, Cysteine
Abstract

Apoptosis is a regulated form of cell death essential to the removal of unwanted cells. At its core, a family of cysteine peptidases named caspases cleave key proteins allowing cell death to occur. To do so, each caspase catalytic pocket recognizes preferred amino acid sequences resulting in proteolysis, but some also use exosites to select and cleave important proteins efficaciously. Such exosites have been found in a few caspases, notably caspase-7 that has a lysine patch (K38KKK) that binds RNA, which acts as a bridge to RNA-binding proteins favoring proximity between the peptidase and its substrates resulting in swifter cleavage. Although caspase-7 interaction with RNA has been identified, in-depth characterization of this interaction is lacking. In this study, using in vitro cleavage assays, we determine that RNA concentration and length affect the cleavage of RNA-binding proteins. Additionally, using binding assays and RNA sequencing, we found that caspase-7 binds RNA molecules regardless of their type, sequence, or structure. Moreover, we demonstrate that the N-terminal peptide of caspase-7 reduces the affinity of the peptidase for RNA, which translates into slower cleavages of RNA-binding proteins. Finally, employing engineered heterodimers, we show that a caspase-7 dimer can use both exosites simultaneously to increase its affinity to RNA because a heterodimer with only one exosite has reduced affinity for RNA and cleavage efficacy. These findings shed light on a mechanism that furthers substrate recognition by caspases and provides potential insight into its regulation during apoptosis.

Published
2021

4. Caspase-7 uses RNA to enhance proteolysis of poly(ADP-ribose) polymerase 1 and other RNA-binding proteins

Author

Alexandre Desroches and Jean-Bernard Denault

Subjects
RNase P, Poly ADP ribose polymerase, Proteolysis, exosite, Amino Acid Motifs, Poly (ADP-Ribose) Polymerase-1, RNA-binding protein, Biochemistry, caspase-7, Substrate Specificity, 03 medical and health sciences, Mice, 0302 clinical medicine, Protein Domains, medicine, Animals, Humans, Binding site, Polymerase, 030304 developmental biology, RNA-BPs, Caspase 7, 0303 health sciences, Multidisciplinary, biology, medicine.diagnostic_test, Chemistry, apoptosis, RNA, RNA-Binding Proteins, Biological Sciences, PNAS Plus, poly(ADP-ribose) polymerase 1, 030220 oncology & carcinogenesis, Nucleic acid, biology.protein, Protein Binding
Abstract

Significance During apoptosis, hundreds of intracellular proteins are cleaved by caspases. In addition to recognizing optimized motifs in the primary structure of its substrates, we show that caspase-7 possesses an exosite to enhance poly(ADP-ribose) polymerase 1 (PARP-1) recognition via the mutual binding of RNA. We also demonstrate that caspase-7 binds RNA and that PARP-1, via 2 RNA-binding domains, likely binds the same RNA molecule, bringing itself near caspase-7 for prompt inactivation. This mechanism is conserved in the mouse ortholog. We have also validated the use of RNA for the efficient cleavage of 6 other RNA-BPs, demonstrating that this distinctive mode of proteolysis enhancement by caspase-7 is an essential feature of substrate recognition during apoptosis., To achieve swift cell demise during apoptosis, caspases cleave essential proteins for cell survival and removal. In addition to the binding of preferred amino acid sequences to its substrate-binding pocket, caspase-7 also uses exosites to select specific substrates. 4 lysine residues (K38KKK) located in the N-terminal domain of caspase-7 form such an exosite and promote the rapid proteolysis of the poly(ADP-ribose) polymerase 1 (PARP-1), but the mechanism of recognition remains mostly unknown. In this study, we show that the overall positive charge of the exosite is the critical feature of this evolutionarily conserved binding site. Additionally, interaction with the caspase-7 exosite involves both the Zn3 and BRCT domains of PARP-1 and is mediated by RNA. Indeed, PARP-1 proteolysis efficacy is sensitive to RNase A and promoted by added RNA. Moreover, using affinity chromatography and gel shift assays, we demonstrate that caspase-7, but not caspase-3 or a caspase-7 with a mutated exosite, binds nucleic acids. Finally, we show that caspase-7 prefers RNA-binding proteins (RNA-BPs) as substrates compared to caspase-3 and that RNA enhances proteolysis by caspase-7 of many of these RNA-BPs. Thus, we have uncovered an unusual way by which caspase-7 selects and cleaves specific substrates.

Published
2019

5. Characterization of Hsp90 Co-Chaperone p23 Cleavage by Caspase-7 Uncovers a Peptidase–Substrate Interaction Involving Intrinsically Disordered Regions

Author

Jean-Bernard Denault, Mikaël Bédard, Cyrielle Martini, and Pierre Lavigne

Subjects
0301 basic medicine, congenital, hereditary, and neonatal diseases and abnormalities, Proteolysis, Biology, Cleavage (embryo), Biochemistry, Caspase 7, Structure-Activity Relationship, 03 medical and health sciences, chemistry.chemical_compound, Protein Domains, Ribose, medicine, Humans, HSP90 Heat-Shock Proteins, Polymerase, Prostaglandin-E Synthases, medicine.diagnostic_test, Circular Dichroism, Phosphoproteins, Hsp90, Co-chaperone, Kinetics, A-site, 030104 developmental biology, chemistry, Mutation, biology.protein, Molecular Chaperones
Abstract

Caspases are cysteinyl peptidases involved in inflammation and apoptosis during which hundreds of proteins are cleaved by executioner caspase-3 and -7. Despite the fact that caspase-3 has a higher catalytic activity, caspase-7 is more proficient at cleaving poly(ADP ribose) polymerase 1 (PARP1) because it uses an exosite within its N-terminal domain (NTD). Here, we demonstrate that molecular determinants also located in the NTD enhance the recognition and proteolysis of the Hsp90 co-chaperone p23. Structure–activity relationship analyses using mutagenesis of the caspase-7 NTD and kinetics show that residues 36–45 of caspase-7, which overlap with residues necessary for efficacious PARP1 cleavage, participate in p23 recognition. We also demonstrate using chimeric and truncated proteins that the caspase-7 NTD binds close to the cleavage site in the C-terminal tail of p23. Moreover, because p23 is cleaved at a site bearing a P4 Pro residue (PEVD142↓G), which is far from the optimal sequence, we tested all res...

Published
2017

6. Caspases rule the intracellular trafficking cartel

Author

Catherine Duclos, Christine Lavoie, and Jean-Bernard Denault

Subjects
0301 basic medicine, Proteolysis, Apoptosis, Biochemistry, 03 medical and health sciences, medicine, Animals, Humans, Nuclear pore, Molecular Biology, Caspase, medicine.diagnostic_test, biology, Cellular process, Intrinsic apoptosis, Cartel, Biological Transport, Cell Biology, Cell biology, 030104 developmental biology, Caspases, Nuclear Pore, biology.protein, Intracellular
Abstract

During apoptosis, caspases feast on several hundreds of cellular proteins to orchestrate rapid cellular demise. Indeed, caspases are known to get a taste of every cellular process in one way or another, activating some, but most often shutting them down. Thus, it is not surprising that caspases proteolyze proteins involved in intracellular trafficking with particularly devastating consequences for this important process. This review article focuses on how caspases target the machinery responsible for smuggling goods within and outside the cell.

Published
2017

7. iRAGE as a novel carboxymethylated peptide that prevents advanced glycation end product-induced apoptosis and endoplasmic reticulum stress in vascular smooth muscle cells

Author

Louis Gendron, Michel Grandbois, Ulrike Froehlich, Jean-Bernard Denault, Jean-Sébastien Maltais, and Elie Simard

Subjects
Glycation End Products, Advanced, 0301 basic medicine, Programmed cell death, Vascular smooth muscle, endocrine system diseases, Myocytes, Smooth Muscle, Receptor for Advanced Glycation End Products, Apoptosis, Biology, Muscle, Smooth, Vascular, Cell Line, RAGE (receptor), 03 medical and health sciences, chemistry.chemical_compound, Glycation, Animals, cardiovascular diseases, Serum Albumin, Caspase 7, Pharmacology, Caspase 3, Lysine, Endoplasmic reticulum, NF-kappa B, nutritional and metabolic diseases, Endoplasmic Reticulum Stress, Rats, Cell biology, 030104 developmental biology, Proto-Oncogene Proteins c-bcl-2, chemistry, Biochemistry, cardiovascular system, Unfolded protein response, Advanced glycation end-product, Peptides, human activities
Abstract

Advanced glycation end-products (AGE) and the receptor for AGE (RAGE) have been linked to numerous diabetic vascular complications. RAGE activation promotes a self-sustaining state of chronic inflammation and has been shown to induce apoptosis in various cell types. Although previous studies in vascular smooth muscle cells (VSMC) showed that RAGE activation increases vascular calcification and interferes with their contractile phenotype, little is known on the potential of RAGE to induce apoptosis in VSMC. Using a combination of apoptotic assays, we showed that RAGE stimulation with its ligand CML-HSA promotes apoptosis of VSMC. The formation of stress granules and the increase in the level of the associated protein HuR point toward RAGE-dependent endoplasmic reticulum (ER) stress, which is proposed as a key contributor of RAGE-induced apoptosis in VSMC as it has been shown to promote cell death via numerous mechanisms, including up-regulation of caspase-9. Chronic NF-κB activation and modulation of Bcl-2 homologs are also suspected to contribute to RAGE-dependent apoptosis in VSMC. With the goal of reducing RAGE signaling and its detrimental impact on VSMC, we designed a RAGE antagonist (iRAGE) derived from the primary amino acid sequence of HSA. The resulting CML peptide was selected for the high glycation frequency of the primary sequence in the native protein in vivo. Pretreatment with iRAGE blocked 69.6% of the increase in NF-κB signaling caused by RAGE activation with CML-HSA after 48h. Preincubation with iRAGE was successful in reducing RAGE-induced apoptosis, as seen through enhanced cell survival by SPR and reduced PARP cleavage. Activation of executioner caspases was 63.5% lower in cells treated with iRAGE before stimulation with CML-HSA. To our knowledge, iRAGE is the first antagonist shown to block AGE-RAGE interaction and we propose the molecule as an initial candidate for drug discovery.

Published
2016

8. Highlight: Frontiers in Proteolysis

Author

Christopher M. Overall, Jean-Bernard Denault, and M. Joanne Lemieux

Subjects
0301 basic medicine, Proteomics, medicine.diagnostic_test, business.industry, Proteolysis, Clinical Biochemistry, Computational biology, Biochemistry, 03 medical and health sciences, 030104 developmental biology, Medicine, Humans, business, Molecular Biology
Published
2018

9. Caspase-mediated proteolysis of the sorting nexin 2 disrupts retromer assembly and potentiates Met/hepatocyte growth factor receptor signaling

Author

Christine Lavoie, Caroline Saucier, Julie Carrier, Catherine Duclos, Jean-Bernard Denault, and Audrey Champagne

Subjects
0301 basic medicine, Cancer Research, biology, Retromer, Endosome, Chemistry, Immunology, Tyrosine phosphorylation, Cell Biology, Article, Transport protein, Cell biology, 03 medical and health sciences, Cellular and Molecular Neuroscience, VPS35, Sorting nexin, chemistry.chemical_compound, 030104 developmental biology, Hepatocyte Growth Factor Receptor, biology.protein, Caspase
Abstract

The unfolding of apoptosis involves the cleavage of hundreds of proteins by the caspase family of cysteinyl peptidases. Among those substrates are proteins involved in intracellular vesicle trafficking with a net outcome of shutting down the crucial processes governing protein transport to organelles and to the plasma membrane. However, because of the intertwining of receptor trafficking and signaling, cleavage of specific proteins may lead to unintended consequences. Here we show that in apoptosis, sorting nexin 1 and 2 (SNX1 and SNX2), two proteins involved in endosomal sorting, are cleaved by initiator caspases and also by executioner caspase-6 in the case of SNX2. Moreover, SNX1 is cleaved at multiple sites, including following glutamate residues. Cleavage of SNX2 results in a loss of association with the endosome-to-trans-Golgi network transport protein Vps35 and in a delocalization from endosomes of its associated partner Vps26. We also demonstrate that SNX2 depletion causes an increase in hepatocyte growth factor receptor tyrosine phosphorylation and Erk1/2 signaling in cells. Finally, we show that SNX2 mRNA and protein levels are decreased in colorectal carcinoma and that lower SNX2 gene expression correlates with an increase in cancer patient mortality. Our study reveals the importance to characterize the cleavage fragments produced by caspases of specific death substrates given their potential implication in the mechanism of regulation of physiological (signaling/trafficking) pathways or in the dysfunction leading to pathogenesis.

Published
2017

10. NLRX1 inhibits the early stages of CNS inflammation and prevents the onset of spontaneous autoimmunity

Author

Carles Vilariño-Güell, Jenny P.-Y. Ting, Peter A. Calabresi, Albert Lamontagne, Véronique Blais, Denis Gris, Kenzo Yamamoto, A. Dessa Sadovnick, Subburaj Ilangumaran, Louis Gendron, Samir Jarjoura, Shaimaa Mahmoud, Abdelaziz Amrani, Diwakar Bobbala, Marjan Gharagozloo, Camille Simard, Matthew D. Smith, and Jean-Bernard Denault

Subjects
Central Nervous System, Male, 0301 basic medicine, Macroglial Cells, Encephalomyelitis, Pathology and Laboratory Medicine, medicine.disease_cause, Nervous System, Autoimmunity, White Blood Cells, Spectrum Analysis Techniques, 0302 clinical medicine, Animal Cells, Medicine and Health Sciences, Biology (General), NLRX1, Immune Response, T Cells, General Neuroscience, Experimental autoimmune encephalomyelitis, Neurodegenerative Diseases, Animal Models, Middle Aged, Flow Cytometry, 3. Good health, medicine.anatomical_structure, Neurology, Spinal Cord, Experimental Organism Systems, Spectrophotometry, Codon, Nonsense, Female, Cytophotometry, Cellular Types, Anatomy, medicine.symptom, General Agricultural and Biological Sciences, Research Article, Adult, Multiple Sclerosis, Encephalomyelitis, Autoimmune, Experimental, QH301-705.5, Immune Cells, Immunology, Mutation, Missense, Mouse Models, Glial Cells, Mice, Transgenic, Inflammation, Biology, Research and Analysis Methods, General Biochemistry, Genetics and Molecular Biology, Autoimmune Diseases, Mitochondrial Proteins, Young Adult, 03 medical and health sciences, Signs and Symptoms, Model Organisms, Diagnostic Medicine, medicine, Animals, Humans, Blood Cells, Innate immune system, General Immunology and Microbiology, Multiple sclerosis, Biology and Life Sciences, Cell Biology, medicine.disease, Demyelinating Disorders, Immunity, Innate, Oligodendrocyte, Neuroanatomy, 030104 developmental biology, Astrocytes, Case-Control Studies, Animal Studies, Clinical Immunology, Clinical Medicine, 030217 neurology & neurosurgery, Neuroscience, Demyelinating Diseases
Abstract

Nucleotide-binding, leucine-rich repeat containing X1 (NLRX1) is a mitochondria-located innate immune sensor that inhibits major pro-inflammatory pathways such as type I interferon and nuclear factor-κB signaling. We generated a novel, spontaneous, and rapidly progressing mouse model of multiple sclerosis (MS) by crossing myelin-specific T-cell receptor (TCR) transgenic mice with Nlrx1−/− mice. About half of the resulting progeny developed spontaneous experimental autoimmune encephalomyelitis (spEAE), which was associated with severe demyelination and inflammation in the central nervous system (CNS). Using lymphocyte-deficient mice and a series of adoptive transfer experiments, we demonstrate that genetic susceptibility to EAE lies within the innate immune compartment. We show that NLRX1 inhibits the subclinical stages of microglial activation and prevents the generation of neurotoxic astrocytes that induce neuronal and oligodendrocyte death in vitro. Moreover, we discovered several mutations within NLRX1 that run in MS-affected families. In summary, our findings highlight the importance of NLRX1 in controlling the early stages of CNS inflammation and preventing the onset of spontaneous autoimmunity., NLRX1 is a guardian protein that inhibits the inflammatory response of glial cells within the central nervous system and prevents the onset of a spontaneous multiple sclerosis–like disease in mice. This study uses a novel mouse model to provide mechanistic insights into the neurodegenerative origin of multiple sclerosis.

Published
2019

11. Organ-specific alteration in caspase expression and STK3 proteolysis during the aging process

Author

Jean-Bernard Denault, Mélissa Lessard-Beaudoin, Rona K. Graham, Mélissa Laroche, Amal Loudghi, Erich E. Wanker, Guillaume Grenier, Sean-Patrick Riechers, and Marie-Josée Demers

Subjects
0301 basic medicine, Male, Aging, Proteolysis, Gene Expression, Apoptosis, Protein Serine-Threonine Kinases, Serine-Threonine Kinase 3, Serine, 03 medical and health sciences, medicine, Animals, Threonine, Caspase, medicine.diagnostic_test, biology, Kinase, General Neuroscience, Intrinsic apoptosis, Neurodegeneration, Brain, Neurodegenerative Diseases, medicine.disease, Cell biology, Mice, Inbred C57BL, 030104 developmental biology, Biochemistry, Organ Specificity, Caspases, biology.protein, Neurology (clinical), Geriatrics and Gerontology, Developmental Biology
Abstract

Caspases and their substrates are key mediators of apoptosis and strongly implicated in various physiological processes. As the serine/threonine kinase family is involved in apoptosis and serine/threonine kinase 3 (STK3) is a recently identified caspase-6 substrate, we assessed the expression and cleavage of STK3 in murine peripheral organs and brain regions during the aging process. We also assessed caspase-3, -6, -7, and -8 expression and activity in order to delineate potential mechanism(s) underlying the generation of the STK3 fragments observed and their relation to the apoptotic pathway. We demonstrate for the first time the cleavage of STK3 by caspase-7 and show that STK3 protein levels globally increase throughout the organism with age. In contrast, caspase-3, -6, -7, and -8 expression and activity vary significantly among the different organs analyzed suggesting differential effects of aging on the apoptotic mechanism and/or nonapoptotic functions of caspases throughout the organism. These results further our understanding of the role of caspases and their substrates in the normal aging process and highlight a potential role for STK3 in neurodegeneration.

Published
2016

12. Age-dependent differential expression of death-associated protein 6 (Daxx) in various peripheral tissues and different brain regions of C57BL/6 male mice

Author

Catherine Duclos, Erich E. Wanker, Mélissa Lessard-Beaudoin, Sean-Patrick Riechers, Jean-Bernard Denault, Marie-Josée Demers, Guillaume Grenier, Mélissa Laroche, and Rona K. Graham

Subjects
0301 basic medicine, Male, Pathology, medicine.medical_specialty, Cell type, Cerebellum, Aging, medicine.disease_cause, 03 medical and health sciences, Mice, Death-associated protein 6, Transcription (biology), medicine, Animals, Caspase, biology, Intracellular Signaling Peptides and Proteins, Brain, Gene Expression Regulation, Developmental, Nuclear Proteins, Cell biology, Mice, Inbred C57BL, Viscera, 030104 developmental biology, medicine.anatomical_structure, Apoptosis, Organ Specificity, biology.protein, Geriatrics and Gerontology, Carcinogenesis, Carrier Proteins, Gerontology, Developmental biology, Co-Repressor Proteins, Molecular Chaperones
Abstract

Death-associated protein 6 (DAXX) is a ubiquitous protein implicated in various cellular processes such as apoptosis, tumorigenesis, development and transcription. The role of DAXX is however ambiguous and many contradictory results regarding its function in apoptosis upon various cellular stresses are described in the literature. In order to have a better understanding of the role of DAXX throughout the entire organism under physiological stress conditions, we have characterized the mRNA levels, protein expression and the proteolytic processing of DAXX in the normal aging process in peripheral organs and brain regions in C57BL/6 male mice. Overall, Daxx mRNA expression decreases with aging in the liver, kidney, heart, cortex and cerebellum. In contrast, an increase is observed in the striatum. The protein expression of DAXX and of its proteolytic fragments increases with aging in the kidney, heart and cortex. In liver and spleen, no changes are observed while in the striatum and cerebellum, certain forms increase and others decrease with age, suggesting that the functions of DAXX may be cell type dependent. This study provides important details regarding the expression and post-translational modifications of DAXX in aging in the entire organism and provides reference data for the deregulation observed in age-associated diseases.

Published
2016

13. Caspase Family

Author

Alexandre Desroches, Dave Boucher, and Jean-Bernard Denault

Published
2016

14. Label-free monitoring of apoptosis by surface plasmon resonance detection of morphological changes

Author

Jean-Sébastien Maltais, Louis Gendron, Michel Grandbois, and Jean-Bernard Denault

Subjects
Pharmacology, Cancer Research, TUNEL assay, biology, Biochemistry (medical), Clinical Biochemistry, Pharmaceutical Science, Apoptosis, Cell Biology, Surface Plasmon Resonance, Cell morphology, biology.organism_classification, Cell biology, TNF-Related Apoptosis-Inducing Ligand, HeLa, Annexin, biology.protein, Humans, Annexin A5, Surface plasmon resonance, Fragmentation (cell biology), Cell Shape, Cells, Cultured, Caspase
Abstract

Apoptosis can be routinely characterized using biomolecular markers such as in the TUNEL and the annexin V assays or by using fluorescent caspase substrates. Apoptosis can also be semi-quantitatively characterized using microscopy, which targets morphological features such as cell rounding, nuclear condensation and fragmentation as well as cell membrane blebbing. This label-free approach provides a limited resolution for the evolution of these events in time and relies heavily on subjective identification of the morphological features. Here we propose a label-free assay based on surface plasmon resonance (SPR) detection of minute morphology changes occurring as a result of apoptosis induction in an endothelial cell model (EA.hy926). At first, annexin V assays confirmed that our cellular model was responsive to TRAIL over a 12-hour period. Then, we show that SPR allows accurate monitoring of apoptosis by measuring (1) the duration of the latency period during which the apoptotic signal is integrated by the initiator caspases and transmitted to the executioner caspases, (2) the rate of the execution phase in which death substrates are cleaved and morphological changes occur, and (3) the total extent of apoptosis. Using these parameters, we characterized the responses obtained with TRAIL (EA.hy926, HeLa, AD-293) and the anti-Fas antibody (HeLa) for the extrinsic pathways and UV exposure (HeLa) for the intrinsic pathways. By comparing the SPR time-course of apoptosis with phase contrast micrographs, we demonstrate that the cell morphological hallmarks of apoptosis are the major contributors to the SPR signal. Altogether, our results validate the use of SPR as an accurate label-free assay for the real-time monitoring of apoptosis-triggered cell morphological changes.

Published
2012

15. Molecular determinants involved in activation of caspase 7

Author

Marcin Drag, Dave Boucher, Véronique Blais, and Jean-Bernard Denault

Subjects
Proteolysis, Caspase 2, Biophysics, Caspase 3, Caspase 8, Biochemistry, Caspase 7, Article, medicine, Humans, Amino Acid Sequence, Molecular Biology, Cells, Cultured, Caspase, Binding Sites, biology, medicine.diagnostic_test, Intrinsic apoptosis, Cell Biology, Caspases, Initiator, Protein Structure, Tertiary, Cell biology, Kinetics, Apoptosis, Caspases, biology.protein, Dimerization
Abstract

During apoptosis, initiator caspases (8, 9 and 10) activate downstream executioner caspases (3, 6 and 7) by cleaving the IDC (interdomain connector) at two sites. Here, we demonstrate that both activation sites, site 1 and site 2, of caspase 7 are suboptimal for activation by initiator caspases 8 and 9 in cellulo, and in vitro using recombinant proteins and activation kinetics. Indeed, when both sites are replaced with the preferred motifs recognized by either caspase 8 or 9, we found an up to 36-fold improvement in activation. Moreover, cleavage at site 1 is preferred to site 2 because of its location within the IDC, since swapping sites does not lead to a more efficient activation. We also demonstrate the important role of Ile195 of site 1 involved in maintaining a network of contacts that preserves the proper conformation of the active enzyme. Finally, we show that the length of the IDC plays a crucial role in maintaining the necessity of proteolysis for activation. In fact, although we were unable to generate a caspase 7 that does not require proteolysis for activity, shortening the IDC of the initiator caspase 8 by four residues was sufficient to confer a requirement for proteolysis, a key feature of executioner caspases. Altogether, the results demonstrate the critical role of the primary structure of caspase 7's IDC for its activation and proteolytic activity.

Published
2011

16. The Transcription Factors NFAT and CREB Have Different Susceptibilities to the Reduced Ca2+ Responses Caused by the Knock Down of Inositol Trisphosphate Receptor in HEK 293A Cells

Author

Jean-Bernard Denault, Guillaume Arguin, Gaétan Guillemette, Annabelle Z. Caron, and Ghadi Elkoreh

Subjects
Gene isoform, NFATC Transcription Factors, biology, Physiology, Chemistry, HEK 293 cells, NFAT, Inositol trisphosphate receptor, CREB, Molecular biology, HEK293 Cells, biology.protein, Humans, Inositol 1,4,5-Trisphosphate Receptors, Calcium, RNA Interference, RNA, Small Interfering, Cyclic AMP Response Element-Binding Protein, Transcription factor, Intracellular
Abstract

Background/Aims: The inositol 1,4,5-trisphosphate receptor (IP3R), a ligand-gated Ca2+ channel, plays an important role in the control of intracellular Ca2+. Three isoforms of IP3R have been identified and most cell types express different proportions of these isoforms. The purpose of this study was to investigate how IP3R signalling is involved in the activation of the Ca2+-sensitive transcription factors NFAT and CREB. Methods: Each IP3R isoform expressed in HEK 293A cells was knocked down using selective siRNA. Free intracellular Ca2+ was monitored spectrofluometrically. NFAT and CREB activities were measured with luciferase reporter constructs. Results: IP3R-2-knocked down HEK 293A cells showed a deficient CCh-induced Ca2+ response that could be rescued by co-stimulation with VIP, a cAMP increasing agonist. NFAT transcriptional activity, but not CREB transcriptional activity, was significantly reduced in IP3R-2-knocked down HEK 293A cells. Overexpression of IP3R-1 could fully compensate for IP3R-2 knock down to mobilize Ca2+ and to activate NFAT. Conclusion: Our results show that the knock down of IP3R-2 significantly reduced the intracellular Ca2+ response of HEK 293 cells. This reduced Ca2+ response did not affect the activation of CREB but significantly decreased the activation of NFAT, suggesting that the Ca2+ signals required for the activation of NFAT are stronger than those required for the activation of CREB.

Published
2010

17. Identification of Early Intermediates of Caspase Activation Using Selective Inhibitors and Activity-Based Probes

Author

Kelly B. Sexton, Amir M. Sadaghiani, Matthew Bogyo, Jean-Bernard Denault, Guy S. Salvesen, Alicia B. Berger, Martin D. Witte, and Stratingh Institute of Chemistry

Subjects
Cell Extracts, Proteases, Proteome, Caspase 2, Apoptosis, Cysteine Proteinase Inhibitors, Cleavage (embryo), Models, Biological, Substrate Specificity, Jurkat Cells, Zymogen, Humans, Molecular Biology, Caspase, Cells, Cultured, Binding Sites, biology, Intrinsic apoptosis, Cell Biology, Caspase Inhibitors, Recombinant Proteins, Cell biology, Enzyme Activation, Kinetics, Biochemistry, Caspases, Molecular Probes, biology.protein, Cysteine
Abstract

Caspases are cysteine proteases that are key effectors in apoptotic cell death. Currently, there is a lack of tools that can be used to monitor the regulation of specific caspases in the context of distinct apoptotic programs. We describe the development of highly selective inhibitors and active site probes and their applications to directly monitor executioner (caspase-3 and -7) and initiator (caspase-8 and -9) caspase activity. Specifically, these reagents were used to dissect the kinetics of caspase activation upon stimulation of apoptosis in cell-free extracts and intact cells. These studies identified a full-length caspase-7 intermediate that becomes catalytically activated early in the pathway and whose further processing is mediated by mature executioner caspases rather than initiator caspases. This form also shows distinct inhibitor sensitivity compared to processed caspase-7. Our data suggest that caspase-7 activation proceeds through a previously uncharacterized intermediate that is formed without cleavage of the intact zymogen.

Published
2006
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18. Yersinia Phosphatase Induces Mitochondrially Dependent Apoptosis of T Cells

Author

Jean-Bernard Denault, Shane Bruckner, Souad Rhamouni, Andres Alonso, Tomas Mustelin, Lutz Tautz, Guy S. Salvesen, and Barbara Becattini

Subjects
Programmed cell death, Yersinia pestis, T-Lymphocytes, bcl-X Protein, Apoptosis, Cell Separation, DNA Fragmentation, Protein tyrosine phosphatase, Transfection, Biochemistry, Jurkat cells, Jurkat Cells, Immune system, Humans, Annexin A5, Phosphorylation, Molecular Biology, Caspase, Cell Death, biology, Cell Biology, Flow Cytometry, Molecular biology, Mitochondria, Nucleosomes, Cell biology, Enzyme Activation, Proto-Oncogene Proteins c-bcl-2, Caspases, Mutagenesis, Site-Directed, biology.protein, Protein Tyrosine Phosphatases, Signal transduction, Tyrosine kinase, Bacterial Outer Membrane Proteins, HeLa Cells, Signal Transduction
Abstract

To evade the immune system, the etiologic agent of plague, Yersinia pestis, injects an exceptionally active tyrosine phosphatase called YopH into host cells using a type III secretion system. We recently reported that YopH acutely inhibits T cell antigen receptor signaling by dephosphorylating the Lck tyrosine kinase. Here, we show that prolonged presence of YopH in primary T cells or Jurkat T leukemia cells causes apoptosis, detected by annexin V binding, mitochondrial breakdown, caspase activation, and internucleosomal fragmentation. YopH also causes cell death when expressed in HeLa cells, and this cell death was inhibited by YopH-specific small molecule inhibitors. Cell death induced by YopH was also prevented by caspase inhibition or co-expression of Bcl-xL. We conclude that YopH not only paralyzes T cells acutely, but also ensures that the cells will not recover to induce a protective immune response but instead undergo mitochondrially regulated programmed cell death.

Published
2005

19. Activation of caspases-8 and -10 by FLIPL

Author

Daniel P. Sutherlin, Guy S. Salvesen, Jean-Bernard Denault, Kelly M. Boatright, and Cristina Deis

Subjects
chemistry.chemical_classification, biology, Chemistry, Activator (genetics), Ripoptosome, Cell Biology, Caspase 8, Biochemistry, Cell biology, Enzyme, Zymogen, biology.protein, FADD, Molecular Biology, Caspase, Death domain
Abstract

The first step in caspase activation is transition of the latent zymogen to an active form. For the initiator caspases, this occurs through dimerization of monomeric zymogens at an activating complex. Recent studies have suggested that FLIPL [FLICE-like inhibitory protein, long form; FLICE is FADD (Fas-associated death domain protein)-like interleukin-1β-converting enzyme], previously thought to act solely as an inhibitor of caspase-8 activation, can under certain circumstances function to enhance caspase activation. Using an in vitro induced-proximity assay, we demonstrate that activation of caspases-8 and -10 occurs independently of cleavage of either the caspase or FLIPL. FLIPL activates caspase-8 by forming heterodimeric enzyme molecules with substrate specificity and catalytic activity indistinguishable from those of caspase-8 homodimers. Significantly, the barrier for heterodimer formation is lower than that for homodimer formation, suggesting that FLIPL is a more potent activator of caspase-8 than is caspase-8 itself.

Published
2004

20. Human Caspase-7 Activity and Regulation by Its N-terminal Peptide

Author

Guy S. Salvesen and Jean-Bernard Denault

Subjects
DNA, Complementary, Time Factors, Recombinant Fusion Proteins, Immunoblotting, Molecular Sequence Data, Apoptosis, DNA Fragmentation, Plasma protein binding, Transfection, Biochemistry, Caspase 7, Cell Line, Substrate Specificity, Cytosol, In vivo, Catalytic Domain, Zymogen, Animals, Humans, Amino Acid Sequence, Molecular Biology, Peptide sequence, Caspase, Cell Nucleus, Sequence Homology, Amino Acid, biology, Caspase 3, Cell Biology, Recombinant Proteins, Protein Structure, Tertiary, Cell biology, Kinetics, Microscopy, Fluorescence, Caspases, Zymogen activation, COS Cells, Mutation, biology.protein, Electrophoresis, Polyacrylamide Gel, Ectopic expression, Peptides, Dimerization, Protein Binding
Abstract

Central to the execution phase of apoptosis are the two closely related caspase-3 and -7. They share common substrate specificity and structure, but differ completely in the sequence of their respective N-terminal regions including their N-peptides, a 23-28 residue segment that are removed during zymogen activation. We show that the N-peptide of caspase-7 plays no role in the fundamental activation or properties of the active protease in vitro. However, the N-peptide modifies the properties of caspase-7 in vivo. In ectopic expression experiments, caspase-7 constructs with no N-peptide are far more lethal than constructs that have an uncleavable peptide. Moreover, the N-peptide of caspase-7 must be removed before efficient activation of the zymogen can occur in vivo. These disparate requirements for the N-peptide argue that it serves to physically sequester the caspase-7 zymogen in a cytosolic location that prevents access by upstream activators (caspase-8, -9, and -10). The N-peptide must first be removed, probably by caspase-3, before efficient conversion and activation of the zymogen can occur in vivo.

Published
2003

21. Caspases: Keys in the Ignition of Cell Death

Author

Guy S. Salvesen and Jean-Bernard Denault

Subjects
Models, Molecular, Programmed cell death, biology, Chemistry, Apoptosis, General Medicine, General Chemistry, Protein Structure, Tertiary, law.invention, Cell biology, Ignition system, law, Caspases, biology.protein, Animals, Humans, Enzyme Inhibitors, Signal transduction, Caspase, Signal Transduction
Published
2002

22. Caspases play in traffic

Author

Christine Lavoie, Julie Carrier, Jean-Bernard Denault, Catherine Duclos, Audrey Champagne, and Caroline Saucier

Subjects
0301 basic medicine, Cancer Research, biology, Chemistry, business.industry, Immunology, News and Commentary, Cell Biology, Computational biology, 03 medical and health sciences, Cellular and Molecular Neuroscience, 030104 developmental biology, Text mining, biology.protein, business, Caspase
Published
2017

23. General in vitro caspase assay procedures

Author

Dave, Boucher, Catherine, Duclos, and Jean-Bernard, Denault

Subjects
Enzyme Activation, Caspases, Humans, Apoptosis, Caspase Inhibitors, Molecular Biology, Recombinant Proteins, Substrate Specificity
Abstract

One of the most valuable tools that have been developed for the study of apoptosis is the availability of recombinant active caspases. The determination of caspase substrate preference, the design of sensitive substrates and potent inhibitors, the resolution of caspase structures, the elucidation of their activation mechanisms, and the identification of their substrates were made possible by the availability of sufficient amounts of enzymatically pure caspases. The current chapter describes at length the expression, purification, and basic enzymatic characterization of apoptotic caspases.

Published
2014

24. General In Vitro Caspase Assay Procedures

Author

Catherine Duclos, Dave Boucher, and Jean-Bernard Denault

Subjects
chemistry.chemical_classification, biology, Substrate (chemistry), In vitro, Enzymatic Assays, law.invention, Cell biology, Enzyme, chemistry, Biochemistry, Apoptosis, law, biology.protein, Recombinant DNA, Caspase
Abstract

One of the most valuable tools that have been developed for the study of apoptosis is the availability of recombinant active caspases. The determination of caspase substrate preference, the design of sensitive substrates and potent inhibitors, the resolution of caspase structures, the elucidation of their activation mechanisms, and the identification of their substrates were made possible by the availability of sufficient amounts of enzymatically pure caspases. The current chapter describes at length the expression, purification, and basic enzymatic characterization of apoptotic caspases.

Published
2014

25. The Contribution of Arginine Residues within the P6–P1 Region of α1-Antitrypsin to Its Reaction with Furin

Author

Pierre Lavigne, Lyne Bissonnette, Erick K. Dufour, Paul C.R. Hopkins, Jean-Bernard Denault, and Richard Leduc

Subjects
Models, Molecular, Time Factors, Stereochemistry, Proteolysis, Immunoblotting, Molecular Sequence Data, Mutant, Context (language use), Arginine, Biochemistry, Protein structure, Leucine, medicine, Amino Acid Sequence, Subtilisins, Binding site, Molecular Biology, Furin, Peptide sequence, Binding Sites, Sequence Homology, Amino Acid, medicine.diagnostic_test, biology, Chemistry, Wild type, Sodium Dodecyl Sulfate, Cell Biology, Recombinant Proteins, Protein Structure, Tertiary, Kinetics, Models, Chemical, alpha 1-Antitrypsin, biology.protein, Electrophoresis, Polyacrylamide Gel, Protein Binding
Abstract

A series of mutants incorporating furin recognition sequences within the P6-P1 region of the reactive site loop of alpha(1)-antitrypsin were constructed. Variants containing different combinations of basic residues in the P1, P2, P4, and P6 positions replacing the wild type (P6)LEAIPM(P1) sequence were evaluated for their capacity to establish SDS-resistant complexes with furin, to affect association rate constants (k(ass) and k'(ass)), or to inhibit furin-dependent proteolysis of a model precursor in vivo. Each variant abolished processing of pro-von Willebrand factor in transfected hEK293 cells. The k(ass) of all variants were found to be similar (1.1-1.7 x 10(6) m(-1) s(-1)) except for one mutant, RERIRR, which had a k(ass) of 3.3 x 10(5) m(-1) s(-1). However, the stoichiometry of inhibition varied with values ranging from 2.9 to >24, indicating rapid formation of the acyl-enzyme intermediate (high k'(ass)). Moreover, those variants having high stoichiometry of inhibition values were accompanied by the rapid formation of cleaved forms of the inhibitors. The data suggest that the rate of conversion of the acyl-enzyme (EI') into the highly stable complex (EI*) was affected by replacement of specific residues within the reactive site loop. Taken together, the results reveal how furin recognition sequences within the context of the biochemical properties of serpins will play a role in the capacity of the protein to follow either the inhibitory or the substrate pathway.

Published
2001

26. Ligand-Induced Signaling in the Absence of Furin Processing of Notch1

Author

Gay Bush, Guy diSibio, Alison Miyamoto, Jean-Bernard Denault, Richard Leduc, and Gerry Weinmaster

Subjects
Cell signaling, Notch, Notch signaling pathway, Receptors, Cell Surface, Ligands, Transfection, Polymerase Chain Reaction, Cell Line, Mice, L Cells, Cell surface receptor, Morphogenesis, Animals, transmembrane receptor, Subtilisins, Receptor, Notch1, Muscle, Skeletal, Furin, Molecular Biology, biology, Membrane Proteins, Cell Differentiation, Cell Biology, Ligand (biochemistry), Recombinant Proteins, Rats, Cell biology, Notch proteins, Hes3 signaling axis, proteolytic processing, Protein Biosynthesis, embryonic structures, Mutagenesis, Site-Directed, biology.protein, Signal transduction, signaling, Dimerization, Signal Transduction, Transcription Factors, Developmental Biology
Abstract

Notch is a conserved cell surface receptor that is activated through direct contact with neighboring ligand-expressing cells. The primary 300-kDa translation product of the Notch1 gene (p300) is cleaved by a furin-like convertase to generate a heterodimeric, cell-surface receptor composed of 180- (p180) and 120- (p120) kDa polypeptides. Heterodimeric Notch is thought to be the only form of the receptor which is both present on the cell surface and able to generate an intracellular signal in response to ligand. Consistent with previous reports, we found that disruption of furin processing of Notch1, either by coexpression of a furin inhibitor or by mutation of furin target sequences within Notch1 itself, perturbed ligand-dependent signaling through the well-characterized mediator of Notch signal transduction, CSL (CBF1, Su(H), and LAG-1). Yet contrary to these reports, we could detect the full-length p300 Notch1 product on the cell surface. Moreover, this uncleaved form of Notch1 could suppress the differentiation of C2C12 myoblasts in response to ligand. Taken together, these data support our previous studies characterizing a CSL-independent Notch signaling pathway and identify this uncleaved isoform of Notch as a potential mediator of this pathway. Our results suggest a novel paradigm in signal transduction, one in which two isoforms of the same cell-surface receptor could mediate two distinct signaling pathways in response to ligand.

Published
2001
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27. Comparative Characterization of Two Forms of Recombinant Human SPC1 Secreted from Schneider 2 Cells

Author

Robert Day, Richard Leduc, Claude Lazure, and Jean-Bernard Denault

Subjects
PNGase F, Glycosylation, Transfection, Amidohydrolases, Cell Line, law.invention, Serine, Sequence Analysis, Protein, law, Enzyme Stability, Animals, Humans, Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase, Protein Isoforms, Cysteine, Subtilisins, Integral membrane protein, Furin, chemistry.chemical_classification, biology, Schneider 2 cells, Membrane Proteins, Hydrogen-Ion Concentration, Recombinant Proteins, Protein Structure, Tertiary, Kinetics, Drosophila melanogaster, Enzyme, Biochemistry, chemistry, Zymogen activation, biology.protein, Recombinant DNA, Calcium, Protein Processing, Post-Translational, Biotechnology
Abstract

SPC1 (furin/PACE), an enzyme belonging to the S8 group of serine endoproteases, is a type I integral membrane protein that catalyzes the processing of a multitude of precursor proteins. We report here the use of transfected Drosophila melanogaster Schneider 2 cells to produce milligram amounts of two forms of recombinant human SPC1. In order to investigate the role of the cysteine-rich region (CRR) of SPC1, we compared the biochemical and enzymatic properties of hSPC1/714 that has the C-terminal tail and transmembrane region of the native enzyme removed with that of hSPC1/585 which had, in addition, the CRR deleted. Two stable cell lines were established. The S2-hSPC1/714 line secreted a major form of apparent molecular weight of 83 kDa and a minor form of 80 kDa whereas the S2-hSPC1/585 line secreted a single 59-kDa protein. PNGase F treatment of the different forms demonstrated that the enzymes were glycosylated. Automated NH 2 -terminal sequencing revealed that all purified forms resulted from processing at the expected zymogen activation site. Removal of the CRR resulted in a broadening of the enzyme's pH range, a shift of K 0.5 for Ca 2+ , and a shorter enzymatic half-life when compared to the longer form, which suggest that the CRR of hSPC1 may help in stabilizing the enzyme's proteolytic activity. The use of this high-level expression system will meet the demand for material necessary to perform biochemical and structural studies that are needed to further our understanding of this and other SPCs at the molecular level.

Published
2000

28. PACE4: a subtilisin-like endoprotease with unique properties

Author

An Zhou, Richard C. Johnson, Jean-Bernard Denault, Richard Leduc, Richard E. Mains, and Carla A. Berard

Subjects
beta-Lipotropin, DNA, Complementary, Blotting, Western, Gene Expression, Prohormone convertase, Biology, Transfection, Biochemistry, chemistry.chemical_compound, Anterior pituitary, medicine, Humans, Secretion, Subtilisins, Peptide Biosynthesis, Cloning, Molecular, Enzyme Inhibitors, Protein precursor, Molecular Biology, Furin, Cells, Cultured, Secretory pathway, Serine Endopeptidases, beta-Endorphin, Leupeptin, Cell Biology, Endonucleases, Immunohistochemistry, medicine.anatomical_structure, chemistry, Pituitary Gland, biology.protein, Electrophoresis, Polyacrylamide Gel, Proprotein Convertases, Protein Processing, Post-Translational, Research Article
Abstract

PACE4 is one of the neuroendocrine-specific mammalian subtilisin-related endoproteases believed to function in the secretory pathway. The biosynthesis and secretion of PACE4 have been studied using transfected neuroendocrine and fibroblast cell lines, as well as primary pituitary cultures. ProPACE4 (approx. 106 kDa) is cleaved intracellularly before secretion of PACE4 (approx. 97 kDa); the N-terminal propeptide cleavage is accelerated in a truncated form of PACE4 lacking the Cys-rich C-terminal region (PACE4s). Neither PACE4 nor PACE4s is stored in regulated neuroendocrine secretory granules, whereas pro-opiomelanocortin-derived peptides and prohormone convertase 1 enter the regulated secretory pathway efficiently. The relatively slow cleavage of the proregion of proPACE4 in primary anterior pituitary cells, followed by rapid secretion of PACE4, is similar to the results for proPACE4 in transfected cell lines. The enzyme activity of PACE4 is distinct from furin and prohormone convertases, both in the marked sensitivity of PACE4 to inhibition by leupeptin and the relative insensitivity of PACE4 to inhibition by Ca2+ chelators and dithiothreitol; PACE4 is not inhibited by the α1-antitrypsin Portland variant that is very potent at inhibiting furin. The unique biosynthetic and enzymic patterns seen for PACE4 suggest a role for this neuroendocrine-specific subtilisin-like endoprotease outside the pathway for peptide biosynthesis.

Published
1997

29. Furin/PACE/SPC1: A convertase involved in exocytic and endocytic processing of precursor proteins

Author

Jean-Bernard Denault and Richard Leduc

Subjects
Saccharomyces cerevisiae Proteins, Molecular Sequence Data, Endocytic cycle, Biophysics, Saccharomyces cerevisiae, Biology, Biochemistry, Exocytosis, Substrate Specificity, Gene product, Exocytic pathway, Structural Biology, Genetics, Animals, Humans, Amino Acid Sequence, Subtilisins, Protein Precursors, Molecular Biology, Furin, Convertase, chemistry.chemical_classification, Proteolytic enzymes, Membrane Proteins, Cell Biology, PACE, Endocytosis, Yeast, Cell biology, Enzyme, chemistry, SPCI, Calcium-Calmodulin-Dependent Protein Kinases, biology.protein, Kexin, Precursor processing, Proprotein Convertases, Schizosaccharomyces pombe Proteins, Mitogen-Activated Protein Kinases, Endocytic pathway, Protein Processing, Post-Translational
Abstract

One of the most exciting breakthroughs of the 90's in the fields of biochemistry, cell biology and neuroendocrinology is the identification of a novel family of proteolytic enzymes called mammalian subtilisin-like convertases. This family is comprised so far of seven distinct endoproteases responsible for the proteolytic excision of biologically active polypeptides from inactive precursor proteins. Six years after the initial observation of a structural conservation between a characterized yeast enzyme (kexin) and a human gene product (furin), it is now well accepted that one of these convertases, furin, has the enzymatic capabilities to efficiently and correctly process a great variety of precursors. Furin's ability to cleave precursors within both the exocytic and endocytic pathways will require sustained efforts in order to delineate all of its physiological roles.

Published
1996

30. Inhibition of Convertase-Related Processing of Proendothelin-1

Author

Jean-Bernard Denault, Tomoh Masaki, Pedro D'Orléans-Juste, and Richard Leduc

Subjects
Pharmacology, chemistry.chemical_classification, biology, Peptide, Cleavage (embryo), Endothelin 1, In vitro, Serine, Enzyme, Biochemistry, chemistry, In vivo, biology.protein, Cardiology and Cardiovascular Medicine, Furin
Abstract

The biologically inactive precursor proendothelin-1 (proET-1) is initially processed intracellularly to the intermediate big endothelin peptide (big ET-1) before its conversion to the endothelin-1 (ET-1) peptide by the endothelin-converting enzyme (ECE). We recently demonstrated that purified furin, a calcium-dependent serine endoprotease belonging to the family of mammalian convertases, cleaved proET-I in vitro and hence produced the physiologically relevant big ET-1 peptide. Therefore, furin becomes a candidate proET-1-cleaving enzyme responsible for the initial biosynthetic processing steps of this precursor. In this study we examined the inhibitory properties of two convertase inhibitors, i.e., the decanoyl-Arg-Val-Lys-Arg-chloromethylketone (dec-RVKR-cmk) and the α 1 -antitrypsin Portland (AT-PDX) on proET-I processing. When purified furin or PACE4, another mammalian convertase, was incubated in the presence of dec-RVKR-cmk, proET-I processing was completely abolished. In the presence of purified AT-PDX, furin-related cleavage of proET-I was abolished but not PACE4 processing. In addition, incubation of endothelial cells with dec-RVKR-cmk inhibited production of ET-1 but did not significantly alter the levels of the prostanoid PGI 2 . These results indicate that convertase-related processing of proET-I can be inhibited in vitro and in vivo, and that convertase-specific processing may be prevented with AT-PDX.

Published
1995

31. CSK-Homologous Kinase

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

32. Cholecystokinin-2 Receptor

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

33. C-type Lectin-Like Receptor 2

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

34. CD191

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

35. c-K-Ras

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

36. CKIP-1 (CK2 Interacting Protein-1)

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

37. C-type Lectin-Domain Family 1 Member A

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

38. c-FGR

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

39. Chemokine (C-X-C Motif) Ligand 10

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

40. CLEC-2

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

41. cyt-PTPe

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

42. C-Type Mannose Receptor 2

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

43. Cysteine Sulfinic Acid Reductase

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

44. C-type Lectin-Domain Family 1, Member b

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

45. C-type Lectin-Like Receptor 1

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

46. C-type Lectin Domain Family 4 Member E

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

47. CCK(A) Receptor

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

48. c-Src Family of Tyrosine Kinases

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

49. C-type (Calcium-Dependent, Carbohydrate-Recognition Domain) lectin, Superfamily Member 5

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

50. Cek4 (Chicken)

Author

Frans van Roy, Volker Nimmrich, Anton Bespalov, Achim Möller, Hiromitsu Hara, Jacob P. Turowec, Nicole A. St. Denis, David W. Litchfield, Dave Boucher, Jean-Bernard Denault, Keiko Matsuda, Michisuke Yuzaki, Carlos Repeke, Thiago Garlet, Ana Paula Trombone, Gustavo Garlet, Elcia Maria Silveira, Andreia Vieira, Fernando Cunha, Satoshi Kubota, Masaharu Takigawa, Helena Soares, Sofia Nolasco, João Gonçalves, Armand Bensussan, Anne Marie-Cardine, Sumit Deswal, Wolfgang W. A. Schamel, Leopoldo Santos-Argumedo, Gail A. Bishop, Dima A. Decker, Bruce S. Hostager, Maria Elena Bravo-Adame, Monserrat Alba Sandoval-Hernandez, Oscar Arturo Migueles-Lozano, Yvonne Rosenstein, Pauline Johnson, Asanga Samarakoon, Amy E. Saunders, Kenneth W. Harder, David D. Roberts, David R. Soto-Pantoja, Jeff S. Isenberg, Pedro A. Lazo, Ramiro Barcia, Hsin-Jung Wu, Natarajan Muthusamy, Subbarao Bondada, Shoshana Levy, Sudesh Pawaria, Robert J. Binder, Hisao Masai, Dongli Hu, Jill M. Lahti, Bernhard B. Singer, Richard Horuk, Laurence J. Miller, Jean Morisset, Anita R. Mistry, Christopher A. O’Callaghan, Angharad E. Fenton-May, Michael Reschen, Janet A. Willment, Gordon D. Brown, Leonard Rabinow, Scott A. Ness, Carl E. Creutz, Nan Yagishita-Kyo, Masatoshi Inoue, Mio Nonaka, Hiroyuki Okuno, Haruhiko Bito, Masato Okada, Heung-Chin Cheng, Mohammed Iqbal Hossain, Mohd Aizuddin Kamaruddin, Yuh-Ping Chong, Banibrata Sen, Faye M. Johnson, Paula A. Pino, Astrid E. Cardona, Federico Paroni, Kathrin Maedler, and Randy Y. C. Poon

Published
2012

Catalog

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