7 results on '"HEAVY-CHAIN"'
Search Results
2. Molecular mechanism for kinesin-1 direct membrane recognition
- Author
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Zuriñe Antón, Judith Mantell, Jessica A. Cross, Edmund R. R. Moody, Yan Y. Yip, Tom A. Williams, Christopher Williams, Roberto A. Steiner, Derek N. Woolfson, Johannes F. Weijman, Mark P. Dodding, and Matthew P. Crump
- Subjects
TERMINAL DOMAIN ,Biophysics ,Kinesins ,BrisSynBio ,CYTOSKELETON ,macromolecular substances ,Microtubules ,Motor protein ,BCS and TECS CDTs ,03 medical and health sciences ,0302 clinical medicine ,Microtubule ,Lysosome ,Organelle ,medicine ,KINESIN ,Health and Medicine ,CYTOSKELETON, KINESIN, LIGHT-CHAIN, HEAVY-CHAIN, TERMINAL DOMAIN, PROTEIN CARGO BINDING IDENTIFICATION, MEMBRANE ,Cytoskeleton ,PROTEIN CARGO BINDING IDENTIFICATION ,Phylogeny ,Research Articles ,030304 developmental biology ,Adaptor Proteins, Signal Transducing ,0303 health sciences ,Multidisciplinary ,LIGHT-CHAIN ,Chemistry ,Bristol BioDesign Institute ,Alternative splicing ,Signal transducing adaptor protein ,SciAdv r-articles ,HEAVY-CHAIN ,Lipids ,Cell biology ,medicine.anatomical_structure ,Kinesin ,synthetic biology ,MEMBRANE ,030217 neurology & neurosurgery ,Research Article - Abstract
Kinesin-1 uses a membrane-induced curvature-sensitive amphipathic helix to bind directly to cargo membranes., The cargo-binding capabilities of cytoskeletal motor proteins have expanded during evolution through both gene duplication and alternative splicing. For the light chains of the kinesin-1 family of microtubule motors, this has resulted in an array of carboxyl-terminal domain sequences of unknown molecular function. Here, combining phylogenetic analyses with biophysical, biochemical, and cell biology approaches, we identify a highly conserved membrane-induced curvature-sensitive amphipathic helix within this region of a subset of long kinesin light-chain paralogs and splice isoforms. This helix mediates the direct binding of kinesin-1 to lipid membranes. Membrane binding requires specific anionic phospholipids, and it contributes to kinesin-1–dependent lysosome positioning, a canonical activity that, until now, has been attributed exclusively the recognition of organelle-associated cargo adaptor proteins. This leads us to propose a protein-lipid coincidence detection framework for kinesin-1–mediated organelle transport.
- Published
- 2021
3. Direct Sensing of Nutrients via a LAT1-like Transporter in Drosophila Insulin-Producing Cells
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Flore Geillon, Anna B. Ziegler, Gérard Manière, David E. Featherstone, Yael Grosjean, Centre des Sciences du Goût et de l'Alimentation [Dijon] (CSGA), Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique (CNRS), Dendrite Differenciation Group [DZNE - Bonn], German Research Center for Neurodegenerative Diseases - Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), Biological Sciences, University of Illinois [Chicago] (UIC), University of Illinois System-University of Illinois System, Centre National de la Recherche Scientifique Université de Bourgogne Franche-ComtéMuscular Dystrophy Association NIH-National Institute of Neurological Disorders and Stroke R01NS045628, Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-Centre National de la Recherche Scientifique (CNRS), and Grosjean, Yaël
- Subjects
0301 basic medicine ,Amino Acid Transport Systems ,heavy-chain ,medicine.medical_treatment ,Insulins ,amino acid transporter ,0302 clinical medicine ,genetics [Drosophila Proteins] ,cytology [Drosophila melanogaster] ,Glutamate Dehydrogenase ,Hemolymph ,Insulin-Secreting Cells ,metabolism [Drosophila melanogaster] ,Drosophila ,Drosophila insulin-like peptides ,food ,glutamate dehydrogenase ,glycemia ,growth ,insulin-producing cells ,minidiscs ,starvation ,Drosophila Proteins ,Protein Isoforms ,metabolism [Calcium] ,genetics [Insulins] ,genetics [Amino Acid Transport Systems] ,lcsh:QH301-705.5 ,Gene knockdown ,cytology [Larva] ,pancreatic beta-cell ,Brain ,metabolism [Hemolymph] ,secretion ,Drosophila melanogaster ,Biochemistry ,Larva ,Alimentation et Nutrition ,Leucine ,Signal Transduction ,glucose-transport ,genetics [Glutamate Dehydrogenase] ,genetics [Protein Isoforms] ,amino-acids ,metabolism [Drosophila Proteins] ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Nutrient sensing ,metabolism [Larva] ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,metabolism [Amino Acid Transport Systems] ,metabolism [Insulins] ,03 medical and health sciences ,parasitic diseases ,medicine ,Food and Nutrition ,Animals ,ddc:610 ,cytology [Insulin-Secreting Cells] ,cardiovascular diseases ,Amino acid transporter ,Mnd protein, Drosophila ,administration & dosage [Leucine] ,metabolism [Protein Isoforms] ,Ilp5 protein, Drosophila ,cytology [Brain] ,Glutamate dehydrogenase ,Insulin ,Neurosciences ,Glucose transporter ,metabolism [Insulin-Secreting Cells] ,glutamate-dehydrogenase ,l-leucine ,030104 developmental biology ,Gene Expression Regulation ,lcsh:Biology (General) ,metabolism [Brain] ,metabolism [Glutamate Dehydrogenase] ,Neurons and Cognition ,metabolism [Leucine] ,Calcium ,metabolism ,fat-cells ,030217 neurology & neurosurgery - Abstract
Summary Dietary leucine has been suspected to play an important role in insulin release, a hormone that controls satiety and metabolism. The mechanism by which insulin-producing cells (IPCs) sense leucine and regulate insulin secretion is still poorly understood. In Drosophila, insulin-like peptides (DILP2 and DILP5) are produced by brain IPCs and are released in the hemolymph after leucine ingestion. Using Ca2+-imaging and ex vivo cultured larval brains, we demonstrate that IPCs can directly sense extracellular leucine levels via minidiscs (MND), a leucine transporter. MND knockdown in IPCs abolished leucine-dependent changes, including loss of DILP2 and DILP5 in IPC bodies, consistent with the idea that MND is necessary for leucine-dependent DILP release. This, in turn, leads to a strong increase in hemolymph sugar levels and reduced growth. GDH knockdown in IPCs also reduced leucine-dependent DILP release, suggesting that nutrient sensing is coupled to the glutamate dehydrogenase pathway., Graphical Abstract, Highlights • IPCs directly sense extracellular leucine levels via minidiscs (MND) • MND knockdown in IPCs abolishes loss of DILP2 and DILP5 • This leads to a strong increase in hemolymph sugar levels and reduces growth • GDH knockdown in IPCs reduces leucine-dependent DILP release, Manière et al. find that leucine induces the disappearance of two DILPs in Drosophila IPCs. Minidiscs (MND) is the primary leucine sensor, and downregulation has consequences for glycemia and growth. The authors propose that the leucine/MND pathway represents a conserved mechanism for insulin release.
- Published
- 2016
4. Bioinspired Silicification of Silica-Binding Peptide-Silk Protein Chimeras: Comparison of Chemically and Genetically Produced Proteins
- Author
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Carole C. Perry, David J. Belton, David L. Kaplan, Heather A. Currie, Laetitia L S Canabady-Rochelle, Olivier Deschaume, CANABADY-ROCHELLE, Laetitia, Nottingham Trent University, Laboratoire Réactions et Génie des Procédés (LRGP), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Tufts Univ, Dept Biomed Engn, Bioengn & Biotechnol Ctr, Tufts University [Medford], and EPSRC EP/E048439/1 AFOSR NIH 5RO1DE017207-05
- Subjects
[SDV.BIO]Life Sciences [q-bio]/Biotechnology ,Spectrophotometry, Infrared ,Polymers and Plastics ,Peptide ,02 engineering and technology ,SPIDER DRAGLINE SILK ,01 natural sciences ,Spectroscopy, Fourier Transform Infrared ,[CHIM] Chemical Sciences ,Materials Chemistry ,Spider silk ,[INFO.INFO-BT]Computer Science [cs]/Biotechnology ,Peptide sequence ,Protein secondary structure ,ComputingMilieux_MISCELLANEOUS ,chemistry.chemical_classification ,biology ,SECONDARY STRUCTURE ,Nephila clavipes ,Silicon Dioxide ,021001 nanoscience & nanotechnology ,BOMBYX-MORI ,[SDV] Life Sciences [q-bio] ,SILK ,Insect Proteins ,FIBROIN ,0210 nano-technology ,Stereochemistry ,Recombinant Fusion Proteins ,Molecular Sequence Data ,Silk ,Fibroin ,Bioengineering ,010402 general chemistry ,SEQUENCE ,Article ,TRANSFORM INFRARED-SPECTROSCOPY ,Biomaterials ,BIOMATERIAL ,Bombyx mori ,Polymer chemistry ,Animals ,[SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering ,Amino Acid Sequence ,fungi ,NANOCONFINEMENT ,BIOSILICIFICATION ,Bombyx ,biology.organism_classification ,HEAVY-CHAIN ,Peptide Fragments ,0104 chemical sciences ,chemistry - Abstract
International audience; Novel protein chimeras constituted of "silk" and a silica-binding peptide (KSLSRHDHIHHH) were synthesized by genetic or chemical approaches and their influence on silica-silk based chimera composite formation evaluated. Genetic chimeras were constructed from 6 or 15 repeats of the 32 amino acid consensus sequence of Nephila clavipes spider silk ([SGRGGLGGQG AGAAAAAGGA GQGGYGGLGSQG](n)) to which one silica binding peptide was fused at the N terminus. For the chemical chimera, 28 equiv of the silica binding peptide were chemically coupled to natural Bombyx mori silk after modification of tyrosine groups by diazonium coupling and EDC/NHS activation of all acid groups. After silica formation under mild, biomaterial-compatible conditions, the effect of peptide addition on the properties of the silk and chimeric silk-silica composite materials was explored. The composite biomaterial properties could be related to the extent of silica condensation and to the higher number of silica binding sites in the chemical chimera as compared with the genetically derived variants. In all cases, the structure of the protein/chimera in solution dictated the type of composite structure that formed with the silica deposition process having little effect on the secondary structural composition of the silk-based materials. Similarly to our study of genetic silk based chimeras containing the R5 peptide (SSKKSGSYSGSKGSKRRIL), the role of the chimeras (genetic and chemical) used in the present study resided more in aggregation and scaffolding than in the catalysis of condensation. The variables of peptide identity, silk construct (number of consensus repeats or silk source), and approach to synthesis (genetic or chemical) can be used to "tune" the properties of the composite materials formed and is a general approach that can be used to prepare a range of materials for biomedical and sensor-based applications.
- Published
- 2017
5. Differential role of nonmuscle myosin II isoforms during blebbing of MCF-7 cells
- Author
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Siddhartha S. Jana, Kankan Bhattacharyya, Shamik Sen, Sumit K. Dey, Kaushik Sengupta, Shekhar Saha, Raman K. Singh, Shyamtanu Chattoraj, and Alakesh Das
- Subjects
0301 basic medicine ,Gene isoform ,Identification ,Myosin light-chain kinase ,Oscillations ,genetic structures ,Biology ,Microtubules ,03 medical and health sciences ,Cell Movement ,Actin Cortex ,Cell Line, Tumor ,Humans ,Protein Isoforms ,Bleb (cell biology) ,Pseudopodia ,Cytoskeleton ,Molecular Biology ,Actin ,Migration ,Nonmuscle Myosin Type IIB ,Heavy-Chain ,Distinct ,Nonmuscle Myosin Type IIA ,fungi ,Requirements ,Cell Biology ,Articles ,Actin cytoskeleton ,eye diseases ,Cell biology ,Actin Cytoskeleton ,Cell Motility ,030104 developmental biology ,MCF-7 Cells ,Mechanism ,sense organs ,Cell Surface Extensions ,Lamellipodium ,Plasma-Membrane - Abstract
One molecular cue that regulates cellular protrusions such as blebbing and lamellipodia in tumor cells has been less explored than other environmental factors. NM II-A induces blebbing and NM II-C1 induces lamellipodia in tumor cells. NM-II isoforms can change the protrusive activity of a tumor cell., Bleb formation has been correlated with nonmuscle myosin II (NM-II) activity. Whether three isoforms of NM-II (NM-IIA, -IIB and -IIC) have the same or differential roles in bleb formation is not well understood. Here we report that ectopically expressed, GFP-tagged NM-II isoforms exhibit different types of membrane protrusions, such as multiple blebs, lamellipodia, combinations of both, or absence of any such protrusions in MCF-7 cells. Quantification suggests that 50% of NM-IIA-GFP–, 29% of NM-IIB-GFP–, and 19% of NM-IIC1-GFP–expressing MCF-7 cells show multiple bleb formation, compared with 36% of cells expressing GFP alone. Of interest, NM-IIB has an almost 50% lower rate of dissociation from actin filament than NM-IIA and –IIC1 as determined by FRET analysis both at cell and bleb cortices. We induced bleb formation by disruption of the cortex and found that all three NM-II-GFP isoforms can reappear and form filaments but to different degrees in the growing bleb. NM-IIB-GFP can form filaments in blebs in 41% of NM-IIB-GFP–expressing cells, whereas filaments form in only 12 and 3% of cells expressing NM-IIA-GFP and NM-IIC1-GFP, respectively. These studies suggest that NM-II isoforms have differential roles in the bleb life cycle.
- Published
- 2016
6. Monomeric IgA can be produced in planta as efficient as IgG, yet receives different N-glycans
- Author
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Maurice Henquet, Dieu-Linh Nguyen, Jaap Bakker, Aska Goverse, Dirk Bosch, Lotte B. Westerhof, Debbie R. van Raaij, Ruud H. P. Wilbers, Cornelis H. Hokke, and Arjen Schots
- Subjects
Glycosylation ,heavy-chain ,plant ,Plant Science ,N-glycosylation ,Immunoglobulin E ,hybrid immunoglobulin ,Mice ,subcellular-localization ,N-linked glycosylation ,Laboratorium voor Plantenfysiologie ,Laboratorium voor Monoklonale Antistoffen ,biology ,Antibodies, Monoclonal ,Plants, Genetically Modified ,Isotype ,terminal propeptide ,insect cells ,BIOS Applied Metabolic Systems ,Ustekinumab ,Antibody ,Laboratory of Plant Physiology ,IgA ,Biotechnology ,medicine.drug ,Protein Binding ,glycosylation ,medicine.drug_class ,Cell Survival ,monoclonal-antibodies ,recombinant antibodies ,Monoclonal antibody ,Antibodies, Monoclonal, Humanized ,Cell Line ,Antibody Isotype ,Immunoglobulin Idiotypes ,Polysaccharides ,Plant Cells ,Tobacco ,medicine ,Animals ,Humans ,Antigens ,Laboratorium voor Nematologie ,Tumor Necrosis Factor-alpha ,Adalimumab ,Infliximab ,Immunoglobulin A ,Immunoglobulin G ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Immunology ,Proteolysis ,biology.protein ,EPS ,Laboratory of Nematology ,infliximab ,protein ,Agronomy and Crop Science - Abstract
The unique features of IgA, such as the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE, make it a promising antibody isotype for several therapeutic applications. However, in contrast to IgG, reports on plant production of IgA are scarce. We produced IgA1¿ and IgG1¿ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-a, and Ustekinumab, directed against the interleukin-12p40 subunit. We evaluated antibody yield, quality and N-glycosylation. All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. However, IgA1¿-based Adalimumab and Ustekinumab were poorly secreted compared to their IgG counterparts. Infliximab was poorly secreted regardless of isotype backbone. This corresponded with the observation that both IgA1¿- and IgG1¿-based Infliximab were enriched in oligomannose-type N-glycan structures. For IgG1¿-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, ß1,2-xylose and core a1,3-fucose. In contrast, the major N-glycan on the IgA-based antibodies was xylosylated, but lacked core a1,3-fucose and one terminal N-acetylglucosamine. This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein. Our data demonstrate that the antibody isotype may have a profound influence on the type of N-glycan an antibody receives.
- Published
- 2014
7. Purification and quantification of heavy-chain antibodies from the milk of bactrian camels.
- Author
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Yao, Hongqiang, Yu, Siriguleng, Zhang, Min, Li, Yi, Yao, Jirimutu, and Meng, He
- Subjects
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IMMUNOGLOBULINS , *BACTRIAN camel , *SODIUM dodecyl sulfate , *CAMEL milk , *BLOOD sugar , *SALT - Abstract
Camel milk has a unique composition with naturally occurring heavy-chain antibodies (HCAbs), which exert rehabilitating potencies in infection and immunity. To characterize HCAb in camel milk, immunoglobulin G (IgG) was isolated from the milk of Camelus bactrianus by a combination of affinity chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis to purify and size-fractionate protein A and protein G, which were further identified by Western blotting, and were quantified by bicinchoninic acid (BCA) and ELISA. The results indicated that IgG1 fraction contains molecules of 50 kDa heavy chains and 36 kDa light chains. The HCAbs (IgG2 and IgG3 fractions) devoid of light chains, contain heavy chains of 45 kDa and 43 kDa, respectively, the amounts of which were significantly higher than that of the IgG1 in the milk of bactrian camels. Above all, we revealed the considerable amounts of HCAbs in the milk of bactrian camels, and developed a novel method for their purification and quantification. These findings provide the basis for developing potential effects of camel milk and its interface with the dairy industry, as well as future investigations of HCAb and its roles in human health and diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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