14 results on '"Harvey, Peta J."'
Search Results
2. The effect of solvent polarity on the rate of the Mitsunobu esterification reaction.
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Camp, David, Harvey, Peta J., and Jenkins, Ian D.
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SOLVENTS , *POLARITY (Chemistry) , *MITSUNOBU reaction , *ESTERIFICATION , *BENZOIC acid , *SODIUM benzoate - Abstract
The rate of the Mitsunobu esterification reaction of ethanol or isopropanol with benzoic acid was found to be much faster in non-polar solvents. The logarithm of the rate constant was inversely proportional to the solvent polarity, as defined by E T values. Typically, the rate constant for ethyl benzoate formation in THF was 100 times greater than that in MeCN. The presence of either sodium benzoate or excess benzoic acid resulted in a decrease in rate. Each of the main species involved in the Mitsunobu esterification reaction, the alcohol starting material, dialkoxyphosphorane, alkoxyphosphonium salt and ester product, was detected by proton NMR analysis. The possible role of ion pair aggregates or clusters, prior to rate-determining S N 2 attack of carboxylate on the alkoxyphosphonium ion, is discussed. An explanation is provided as to why the yield in the Mitsunobu reaction is often higher in non-polar solvents. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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3. Alternative splicing of RyR1 alters the efficacy of skeletal EC coupling.
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Kimura, Takashi, Lueck, John D., Harvey, Peta J., Pace, Suzy M., Ikemoto, Noriaki, Casarotto, Marco G., Dirksen, Robert T., and Dulhunty, Angela F.
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PROTEIN engineering ,RYANODINE receptors ,MYOTONIA atrophica ,MUSCLE contraction ,CALCIUM channels ,ANTIBIOTIC residues ,GENE expression - Abstract
Abstract: Alternative splicing of ASI residues (Ala
3481 –Gln3485 ) in the skeletal muscle ryanodine receptor (RyR1) is developmentally regulated: the residues are present in adult ASI(+)RyR1, but absent in the juvenile ASI(−)RyR1 which is over-expressed in adult myotonic dystrophy type 1 (DM1). Although this splicing switch may influence RyR1 function in developing muscle and DM1, little is known about the properties of the splice variants. We examined excitation-contraction (EC) coupling and the structure and interactions of the ASI domain (Thr3471 –Gly3500 ) in the splice variants. Depolarisation-dependent Ca2+ release was enhanced by >50% in myotubes expressing ASI(−)RyR1 compared with ASI(+)RyR1, although DHPR L-type currents and SR Ca2+ content were unaltered, while ASI(−)RyR1 channel function was actually depressed. The effect on EC coupling did not depend on changes in ASI domain secondary structure. Probing RyR1 function with peptides possessing the ASI domain sequence indicated that the domain contributes to an inhibitory module in RyR1. The action of the peptide depended on a sequence of basic residues and their alignment in an α-helix adjacent to the ASI splice site. This is the first evidence that the ASI residues contribute to an inhibitory module in RyR1 that influences EC coupling. Implications for development and DM1 are discussed. [Copyright &y& Elsevier]- Published
- 2009
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4. Critical residue properties for potency and selectivity of α-Conotoxin RgIA towards α9α10 nicotinic acetylcholine receptors.
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Huynh, Peter N., Harvey, Peta J., Gajewiak, Joanna, Craik, David J., and Michael McIntosh, J.
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NICOTINIC acetylcholine receptors , *CONOTOXINS , *CHEMICAL properties , *ION channels , *CHRONIC pain - Abstract
The α9α10 nicotinic acetylcholine receptor (nAChR) has been characterized as an effective anti-pain target that functions through a non-opioid mechanism. However, as a pentameric ion channel comprised of two different subunits, the specific targeting of α9α10 nAChRs has proven challenging. Previously the 13-amino-acid peptide, RgIA, was shown to block α9α10 nAChRs with high potency and specificity. This peptide, characterized from the venom of the carnivorous marine snail, Conus regius , produced analgesia in several rodent models of chronic pain. Despite promising pre-clinical data in behavioral assays, the number of specific α9α10 nAChR antagonists remains small and the physiological mechanisms of analgesia remain cryptic. In this study, we implement amino-acid substitutions to definitively characterize the chemical properties of RgIA that contribute to its activity against α9α10 nAChRs. Using this mutational approach, we determined the vital role of biochemical side-chain properties and amino acids in the second loop that are amenable to substitutions to further engineer next-generation analogs for the blockade of α9α10 nAChRs. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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5. Design, synthesis, and mechanism of action of novel μ-conotoxin KIIIA analogues for inhibition of the voltage-gated sodium channel Nav1.7.
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Zitong Zhao, Teng Pan, Shen Chen, Harvey, Peta J., Jinghui Zhang, Xiao Li, Mengke Yang, Linhong Huang, Shoushi Wang, Craik, David J., Tao Jiang, and Rilei Yu
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SODIUM channels , *MOLECULAR dynamics , *PEPTIDES , *SODIUM channel blockers , *PAIN management - Abstract
µ-Conotoxin KIIIA, a selective blocker of sodium channels, has strong inhibitory activity against several Nav isoforms, including Nav1.7, and has potent analgesic effects, but it contains three pairs of disulfide bonds, making structural modification difficult and synthesis complex. To circumvent these difficulties, we designed and synthesized three KIIIA analogues with one disulfide bond deleted. The most active analogue, KIIIA-1, was further analyzed, and its binding pattern to hNav1.7 was determined by molecular dynamics simulations. Guided by the molecular dynamics computational model, we designed and tested 32 second-generation and 6 thirdgeneration analogues of KIIIA-1 on hNav1.7 expressed in HEK293 cells. Several analogues showed significantly improved inhibitory activity on hNav1.7, and the most potent peptide, 37, was approximately 4-fold more potent than the KIIIA Isomer I and 8-fold more potent than the wildtype (WT) KIIIA in inhibiting hNav1.7 current. Intraperitoneally injected 37 exhibited potent in vivo analgesic activity in a formalininduced inflammatory pain model, with activity reaching -350-fold of the positive control drug morphine. Overall, peptide 37 has a simplified disulfide-bond framework and exhibits potent in vivo analgesic effects and has promising potential for development as a pain therapy in the future. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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6. The acyclotide ribe 31 from Rinorea bengalensis has selective cytotoxicity and potent insecticidal properties in Drosophila.
- Author
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Tien T. Dang, Yen-Hua Huang, Ott, Stanislav, Harvey, Peta J., Gilding, Edward K., Tombling, Benjamin J., Lai Y. Chan, Kaas, Quentin, Claridge-Chang, Adam, and Craik, David J.
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DROSOPHILIDAE , *SURFACE plasmon resonance , *DROSOPHILA , *DROSOPHILA melanogaster , *ERYTHROCYTES - Abstract
Cyclotides and acyclic versions of cyclotides (acyclotides) are peptides involved in plant defense. These peptides contain a cystine knot motif formed by three interlocked disulfide bonds, with the main difference between the two classes being the presence or absence of a cyclic backbone, respectively. The insecticidal activity of cyclotides is well documented, but no study to date explores the insecticidal activity of acyclotides. Here, we present the first in vivo evaluation of the insecticidal activity of acyclotides from Rinorea bengalensis on the vinegar fly Drosophila melanogaster. Of a group of structurally comparable acyclotides, ribe 31 showed the most potent toxicity when fed to D. melanogaster. We screened a range of acyclotides and cyclotides and found their toxicity toward human red blood cells was substantially lower than toward insect cells, highlighting their selectivity and potential for use as bioinsecticides. Our confocal microscopy experiments indicated their cytotoxicity is likely mediated via membrane disruption. Furthermore, our surface plasmon resonance studies suggested ribe 31 preferentially binds to membranes containing phospholipids with phosphatidyl-ethanolamine headgroups. Despite having an acyclic backbone, we determined the three-dimensional NMR solution structure of ribe 31 is similar to that of cyclotides. In summary, our results suggest that, with further optimization, ribe 31 could have applications as an insecticide due to its potent in vivo activity against D. melanogaster. More broadly, this work advances the field by demonstrating that acyclotides are more common than previously thought, have potent insecticidal activity, and have the advantage of potentially being more easily manufactured than cyclotides. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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7. The radish defensins RsAFP1 and RsAFP2 act synergistically with caspofungin against Candida albicans biofilms.
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Vriens, Kim, Cools, Tanne L., Harvey, Peta J., Craik, David J., Braem, Annabel, Vleugels, Jozef, Coninck, Barbara De, Cammue, Bruno P.A., and Thevissen, Karin
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CANDIDA albicans , *DEFENSINS , *ANTIFUNGAL agents , *APOPTOSIS , *BIOFILMS , *PHYTOPATHOGENIC microorganisms - Abstract
The radish defensin RsAFP2 was previously characterized as a peptide with potent antifungal activity against several plant pathogenic fungi and human pathogens, including Candida albicans . RsAFP2 induces apoptosis and impairs the yeast-to-hypha transition in C. albicans . As the yeast-to-hypha transition is considered important for progression to mature biofilms, we analyzed the potential antibiofilm activity of recombinant (r)RsAFP2, heterologously expressed in Pichia pastoris , against C. albicans biofilms. We found that rRsAFP2 prevents C. albicans biofilm formation with a BIC-2 ( i.e. , the minimal rRsAFP2 concentration that inhibits biofilm formation by 50% as compared to control treatment) of 1.65 ± 0.40 mg/mL. Moreover, biofilm-specific synergistic effects were observed between rRsAFP2 doses as low as 2.5 μg/mL to 10 μg/mL and the antimycotics caspofungin and amphotericin B, pointing to the potential of RsAFP2 as a novel antibiofilm compound. In addition, we characterized the solution structure of rRsAFP2 and compared it to that of RsAFP1, another defensin present in radish seeds. These peptides have similar amino acid sequences, except for two amino acids, but rRsAFP2 is more potent than RsAFP1 against planktonic and biofilm cultures. Interestingly, as in case of rRsAFP2, also RsAFP1 acts synergistically with caspofungin against C. albicans biofilms in a comparable low dose range as rRsAFP2. A structural comparison of both defensins via NMR analysis revealed that also rRsAFP2 adopts the typical cysteine-stabilized αβ-motif of plant defensins, however, no structural differences were found between these peptides that might result in their differential antifungal/antibiofilm potency. This further suggests that the conserved structure of RsAFP1 and rRsAFP2 bears the potential to synergize with antimycotics against C. albicans biofilms. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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8. Neurotoxic and cytotoxic peptides underlie the painful stings of the tree nettle Urtica ferox.
- Author
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Jing Xie, Robinson, Samuel D., Gilding, Edward K., Jami, Sina, Deuis, Jennifer R., Rehm, Fabian B. H., Yap, Kuok, Ragnarsson, Lotten, Lai Yue Chan, Hamilton, Brett R., Harvey, Peta J., Craik, David J., Vetter, Irina, and Durek, Thomas
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STINGING nettle , *PEPTIDES , *SODIUM channels , *ACTIVATION energy , *CELL membranes , *TOXINS - Abstract
The stinging hairs of plants from the family Urticaceae inject compounds that inflict pain to deter herbivores. The sting of the New Zealand tree nettle (Urtica ferox) is among the most painful of these and can cause systemic symptoms that can even be life-threatening; however, the molecular species effecting this response have not been elucidated. Here we reveal that two classes of peptide toxin are responsible for the symptoms of U. ferox stings: Δ-Uf1a is a cytotoxic thionin that causes pain via disruption of cell membranes, while β/δ-Uf2a defines a new class of neurotoxin that causes pain and systemic symptoms via modulation of voltage-gated sodium (NaV) channels. We demonstrate using whole-cell patch-clamp electrophysiology experiments that β/δ-Uf2a is a potent modulator of human NaV1.5 (EC50: 55 nM), NaV1.6 (EC50: 0.86 nM), and NaV1.7 (EC50: 208 nM), where it shifts the activation threshold to more negative potentials and slows fast inactivation. We further found that both toxin classes are widespread among members of the Urticeae tribe within Urticaceae, suggesting that they are likely to be pain-causing agents underlying the stings of other Urtica species. Comparative analysis of nettles of Urtica, and the recently described pain-causing peptides from nettles of another genus, Dendrocnide, indicates that members of tribe Urticeae have developed a diverse arsenal of pain-causing peptides. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
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9. A Centipede Toxin Family Defines an Ancient Class of CSαβ Defensins.
- Author
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Dash, Thomas S., Shafee, Thomas, Harvey, Peta J., Zhang, Chuchu, Peigneur, Steve, Deuis, Jennifer R., Vetter, Irina, Tytgat, Jan, Anderson, Marilyn A., Craik, David J., Durek, Thomas, and Undheim, Eivind A.B.
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DEFENSINS , *PEPTIDES , *VENOM , *CENTIPEDES , *PROTEIN folding , *PROKARYOTES - Abstract
Summary Disulfide-rich peptides (DRPs) play diverse physiological roles and have emerged as attractive sources of pharmacological tools and drug leads. Here we describe the 3D structure of a centipede venom peptide, U-SLPTX 15 -Sm2a, whose family defines a unique class of one of the most widespread DRP folds known, the cystine-stabilized α/β fold (CSαβ). This class, which we have named the two-disulfide CSαβ fold (2ds-CSαβ), contains only two internal disulfide bonds as opposed to at least three in all other confirmed CSαβ peptides, and constitutes one of the major neurotoxic peptide families in centipede venoms. We show the 2ds-CSαβ is widely distributed outside centipedes and is likely an ancient fold predating the split between prokaryotes and eukaryotes. Our results provide insights into the ancient evolutionary history of a widespread DRP fold and highlight the usefulness of 3D structures as evolutionary tools. Graphical Abstract Highlights • A centipede toxin structure defines a distinct type of the CSαβ defensin fold • This fold is characterized by two disulfides versus at least three in other CSαβ peptides • 2ds-CSαβ peptides are widespread in both eukaryotes and prokaryotes • The 2ds-CSαβ fold probably has an ancient pre-eukaryotic origin Disulfide-rich peptides (DRPs) play many important physiological roles, and can be extremely taxonomically widespread. Here, Dash et al. show that a diverse centipede toxin family belongs to one of the most widespread DRP folds known, the cysteine-stabilized α/β fold, but that it represents a unique, ancient form of this fold. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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10. Molecular determinants of α-conotoxin potency for inhibition of human and rat α6β4 nicotinic acetylcholine receptors.
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Hone, Arik J., Talley, Todd T., Bobango, Janet, Melo, Cesar Huidobro, Hararah, Fuaad, Gajewiak, Joanna, Christensen, Sean, Harvey, Peta J., Craik, David J., and McIntosh, J. Michael
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CONOTOXINS , *NICOTINIC acetylcholine receptors , *DORSAL root ganglia , *LIGAND binding (Biochemistry) , *CRYSTALLOGRAPHY - Abstract
Nicotinic acetylcholine receptors (nAChRs) containing α6 and β4 subunits are expressed by dorsal root ganglion neurons and have been implicated in neuropathic pain. Rodent models are often used to evaluate the efficacy of analgesic compounds, but species differences may affect the activity of some nAChR ligands. A previous candidate α-conotoxin-based therapeutic yielded promising results in rodent models, but failed in human clinical trials, emphasizing the importance of understanding species differences in ligand activity. Here, we show that human and rat α6/α3β4 nAChRs expressed in Xenopus laevis oocytes exhibit differential sensitivity toα-conotoxins. Sequence homology comparisons of human and ratα6β4 nAChR subunits indicated that α6 residues forming the ligand-binding pocket are highly conserved between the two species, but several residues of β4 differed, including a Leu-Gln difference at position 119. X-ray crystallography of α-conotoxin PeIA complexed with the Aplysia californica acetylcholine-binding protein (AChBP) revealed that binding of PeIA orients Pro13 in close proximity to residue 119 of the AChBP complementary subunit. Sitedirected mutagenesis studies revealed that Leu119 of human β4 contributes to higher sensitivity of human α6/α3β4 nAChRs toα-conotoxins, and structure-activity studies indicated that PeIA Pro13 is critical for high potency. Human and rat α6/α3β4 nAChRs displayed differential sensitivities to perturbations of the interaction between PeIA Pro13 and residue 119 of the β4 subunit. These results highlight the potential significance of species differences in α6β4 nAChR pharmacology that should be taken into consideration when evaluating the activity of candidate human therapeutics in rodent models. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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11. Backbone cyclization of analgesic conotoxin GeXIVA facilitates direct folding of the ribbon isomer.
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Xiaosa Wu, Yen-Hua Huang, Kaas, Quentin, Harvey, Peta J., Wang, Conan K., Han-Shen Tae, Adams, David J., and Craik, David J.
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CONOTOXINS , *CHOLINERGIC receptors , *PEPTIDES , *CHEMICAL yield , *DISULFIDES - Abstract
Conotoxin GeXIVA inhibits the α9α10 nicotinic acetylcholine receptor (nAChR) and is analgesic in animal models of pain. α-Conotoxins have four cysteines that can have three possible disulfide connectivities: globular (CysI-CysIII and CysII-CysIV), ribbon (CysI-CysIV and CysII-CysIII), or bead (CysI-CysII and CysIII-CysIV). Native α-conotoxins preferably adopt the globular connectivity, and previous studies of α-conotoxins have focused on the globular isomers as the ribbon and bead isomers typically have lower potency at nAChRs than the globular form. A recent report showed that the bead and ribbon isomers of GeXIVA are more potent than the globular isomer, with low nanomolar half-maximal inhibitory concentrations (IC50). Despite this high potency, the therapeutic potential of GeXIVA is limited, because like most peptides, it is susceptible to proteolytic degradation and is challenging to synthesize in high yield. Here we used backbone cyclization as a strategy to improve the folding yield as well as increase the serum stability of ribbon GeXIVA while preserving activity at the α9α10 nAChR. Specifically, cyclization of ribbon GeXIVA with a two-residue linker maintained the biological activity at the human α9α10 nAChR and improved stability in human serum. Short linkers led to selective formation of the ribbon disulfide isomer without requiring orthogonal protection. Overall, this study highlights the value of backbone cyclization in directing folding, improving yields, and stabilizing conotoxins with therapeutic potential. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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12. Phage display-based discovery of cyclic peptides against the broad spectrum bacterial anti-virulence target CsrA.
- Author
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Jakob, Valentin, Zoller, Ben G.E., Rinkes, Julia, Wu, Yingwen, Kiefer, Alexander F., Hust, Michael, Polten, Saskia, White, Andrew M., Harvey, Peta J., Durek, Thomas, Craik, David J., Siebert, Andreas, Kazmaier, Uli, and Empting, Martin
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CYCLIC peptides , *YERSINIA pseudotuberculosis , *PEPTIDES , *PHARMACEUTICAL chemistry , *PEPTIDE synthesis , *MACROCYCLIC compounds , *BACTERIOPHAGES - Abstract
Small macrocyclic peptides are promising candidates for new anti-infective drugs. To date, such peptides have been poorly studied in the context of anti-virulence targets. Using phage display and a self-designed peptide library, we identified a cyclic heptapeptide that can bind the carbon storage regulator A (CsrA) from Yersinia pseudotuberculosis and displace bound RNA. This disulfide-bridged peptide, showed an IC50 value in the low micromolar range. Upon further characterization, cyclisation was found to be essential for its activity. To increase metabolic stability, a series of disulfide mimetics were designed and a redox-stable 1,4-disubstituted 1,2,3-triazole analogue displayed activity in the double-digit micromolar range. Further experiments revealed that this triazole peptidomimetic is also active against CsrA from Escherichia coli and RsmA from Pseudomonas aeruginosa. This study provides an ideal starting point for medicinal chemistry optimization of this macrocyclic peptide and might pave the way towards broad-acting virulence modulators. [Display omitted] • Screening of self-designed phage library yielded very short macrocyclic peptide hits. • Identification of single-digit micromolar CsrA-RNA interaction inhibitor. • Synthesis of Triazole peptide as redox stable disulfide mimic. • NMR derived solution structure + docking experiments. • Activity on CsrA/RsmA from several species holds promise for broader acting antivirulence agents against Gram-negatives. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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13. Effects of an α-helical ryanodine receptor C-terminal tail peptide on ryanodine receptor activity: Modulation by Homer
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Pouliquin, Pierre, Pace, Suzy M., Curtis, Suzanne M., Harvey, Peta J., Gallant, Esther M., Zorzato, Francesco, Casarotto, Marco G., and Dulhunty, Angela F.
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RYANODINE receptors , *PEPTIDE hormones , *PROTEINS , *CYTOSKELETON - Abstract
Abstract: We have determined the structure of a domain peptide corresponding to the extreme 19 C-terminal residues of the ryanodine receptor Ca2+ release channel. We examined functional interactions between the peptide and the channel, in the absence and in the presence of the regulatory protein Homer. The peptide was partly α-helical and structurally homologous to the C-terminal end of the T1 domain of voltage-gated K+ channels. The peptide (0.1–10μM) inhibited skeletal ryanodine receptor channels when the cytoplasmic Ca2+ concentration was 1μM; but with 10μM cytoplasmic Ca2+, skeletal ryanodine receptors were activated by ≤1.0μM peptide and inhibited by 10μM peptide. Cardiac ryanodine receptors on the other hand were inhibited by all peptide concentrations, at both Ca2+ concentrations. When channels did open in the presence of the peptide, they were more likely to open to substate levels. The inhibition and increased fraction of openings to subconductance levels suggested that the domain peptide might destabilise inter-domain interactions that involve the C-terminal tail. We found that Homer 1b not only interacts with the channels, but reduces the inhibitory action of the C-terminal tail peptide, perhaps by stabilizing inter-domain interactions and preventing their disruption. [Copyright &y& Elsevier]
- Published
- 2006
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14. Backbone cyclization of analgesic conotoxin gexiva facilitates direct folding of the ribbon isomer.
- Author
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Wu, Xiaosa, Huang, Yen-Hua, Kaas, Quentin, Harvey, Peta J., Wang, Conan K., Tae, Han-Shen, Adams, David J., and Craik, David J.
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CONOTOXINS , *ANALGESICS , *ISOMERS , *RING formation (Chemistry) , *CONUS , *MARINE toxins - Published
- 2019
- Full Text
- View/download PDF
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