The Synthesis of Cystine Peptides by Iodine Oxidation of <i>S</i>‐Trityl‐cysteine and <i>S</i>‐Acetamidomethyl‐cysteine Peptides

Kamber, B.; Hartmann, A. A.; Eisler, K.; Riniker, B.; Rink, H; Sieber, Peter; Rittel, W.

doi:10.1002/hlca.19800630418

Cited by 277 publications

(188 citation statements)

References 45 publications

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“…Trypsin was from Promega (Madison, WI). Endothelin-1 was synthesized with Fmoc chemistry as described previously (28)(29)(30)(31).…”

Section: Methodsmentioning

confidence: 99%

Post-translational Modifications of Endothelin Receptor B from Bovine Lungs Analyzed by Mass Spectrometry

Roos¹,

Šoškić²,

Poznanović³

et al. 1998

Journal of Biological Chemistry

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A new mild experimental approach for isolation of peptide membrane receptors and subsequent analysis of post-translational modifications is described. Endothelin receptors A and B were isolated on oligo(dT)-cellulose using N-(⑀-maleimidocaproyloxy)succinimide endothelin coupled to a protected (dA)-30-mer. This allowed a one-step isolation of the receptor from oligo(dT)-cellulose via variation solely of salt concentration. The identity of the receptor was confirmed by direct amino acid sequencing of electroblotted samples or by using antibodies against ET A and ET B receptors. Although it is clear that post-translational modifications of G-protein-coupled receptors are intimately involved in the physiological function of these signal transduction systems, there is as yet relatively little direct evidence for the specific modifications and the relationship of the different modifications to signal transduction processes. These types of processes are not easily amenable to analysis by genomic methods, i.e. there is a need for efficient methods to directly analyze these processes at the proteome level. We report here new, efficient methods for rapid isolation of the endothelin B receptor and for highly sensitive analysis of its post-translational modifications via mass spectrometry.Endothelin, the strongest vasoconstrictor yet known, is a 21-amino acid peptide with physiological effects on cellular development, differentiation, vasoconstriction, and mitogenesis (1, 2). There are 3 different endothelin isoforms, ET-1, 1 ET-2, and ET-3 with different affinity for the two different endothelin receptor subtypes, A and B (3-6). Both receptors are members of the G-protein-coupled receptor superfamily (3-6, 7). The consensus ET receptor topology includes three extracellular domains, three intracellular loops, and a cytoplasmic COOH-terminal tail, separated by seven hydrophobic helical regions thought to span the lipid bilayer. In addition it has been presumed that ET receptors are post-translationally modified by glycosylation of the NH 2 terminus and by phosphorylation and palmitoylation of the cytoplasmic surface. Based on homology with other G-protein-coupled receptors (8 -10), there has been speculation regarding possible structures, functional regions, and sites of post-translational modifications for endothelin receptors (11-16). However, as yet there is very little direct evidence for attributes such as the sites and the roles of glycosylation, palmitoylation, and phosphorylation, the location of the endothelin-binding site and the basis for discrimination among the three different endothelin isoforms.A role of endothelin in disease has recently been demonstrated by the finding that mutated ET B receptor is associated with Hirschsprung's disease (17-21). In addition, mice lacking the ET-1 gene display severe malformation of large blood vessels, stressing the importance of endothelin during development (22). Endothelin has been shown to be a mitogenic agonist in different cell types (23, 24). The signaling pathway by ...

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“…Trypsin was from Promega (Madison, WI). Endothelin-1 was synthesized with Fmoc chemistry as described previously (28)(29)(30)(31).…”

Section: Methodsmentioning

confidence: 99%

Post-translational Modifications of Endothelin Receptor B from Bovine Lungs Analyzed by Mass Spectrometry

Roos¹,

Šoškić²,

Poznanović³

et al. 1998

Journal of Biological Chemistry

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“…Disulfide bridge formation was carried out while peptides were on the solid support. 59,60 Cys residues were deprotected selectively and simultaneously oxidized by adding 70 mg I 2 dissolved in 5 ml 80% (v/v) acetic acid/water to each 100 mg of the peptide resin. I 2 oxidation was allowed to proceed for 8 h. Then the peptide-resin was washed free of I 2 with DMF.…”

Section: Peptide Synthesis Purification and Characterizationmentioning

confidence: 99%

Optical spectroscopic elucidation of β‐turns in disulfide bridged cyclic tetrapeptides

2006

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Vibrational circular dichroism (VCD) spectroscopic features of type II β‐turns were characterized previously, but, criteria for differentiation between β‐turn types had not been established yet. Model tetrapeptides, cyclized through a disulfide bridge, were designed on the basis of previous experimental results and the observed incidence of amino acid residues in the i + 1 and i + 2 positions in β‐turns, to determine the features of VCD spectra of type I and II β‐turns. The results were correlated with electronic circular dichroism (ECD) spectra and VCD spectra calculated from conformational data obtained by molecular dynamics (MD) simulations. All cyclic tetrapeptides yielded VCD signals with a higher frequency negative and a lower frequency positive couplet with negative lobes overlapping. MD simulations confirmed the conformational homogeneity of these peptides in solution. Comparison with ECD spectroscopy, MD, and quantum chemical calculation results suggested that the low frequency component of VCD spectra originating from the tertiary amide vibrations could be used to distinguish between types of β‐turn structures. On the basis of this observation, VCD spectroscopic features of type II and VIII β‐turns and ECD spectroscopic properties of a type VIII β‐turn were suggested. The need for independent experimental as well as theoretical investigations to obtain decisive conformational information was recognized. © 2006 Wiley Periodicals, Inc. Biopolymers 85: 1–11, 2007.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

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“…The peptides were synthesized as described above in the general procedure. Disulfide bonds within these peptides were formed by oxidation of the two Cys[trityl (Trt)] amino acids with 0.1 M iodine/MeOH (Kamber et al, 1980). The respective calculated and experimentally found molecular weight values were 1928.20 and 1928.01 for compound 5 (C*WKTSRTSHYC*PLYQG), and 1896.8519 and 1896.8518 for compound 6 (C*WKTSRTSHYC*PLYPG).…”

Section: Peptide Antagonists With Antiangiogenic Activitymentioning

confidence: 99%

Vimocin and Vidapin, Cyclic KTS Peptides, Are Dual Antagonists ofα₁β₁/α₂β₁Integrins with Antiangiogenic Activity

Momić

Katzehendler

Benny

et al. 2014

J Pharmacol Exp Ther

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Obtustatin and viperistatin, members of the disintegrin protein family, served as lead compounds for the synthesis of linear and cyclic peptides containing the KTS binding motif. The most active linear peptide, a viperistatin analog, indicated the importance of Cys 19 and Cys 29 , as well as the presence of Arg at position 24 for their biologic activity, and was used as the basic sequence for the synthesis of cyclic peptides. Vimocin (compound 6) and vidapin (compound 10) showed a high potency (IC 50 = 0.17 nM) and intermediate efficacy (20 and 40%) in inhibition of adhesion of a 1 /a 2 integrin overexpressor cells to respective collagens. Vimocin was more active in inhibition of the wound healing (53%) and corneal micropocket (17%) vascularization, whereas vidapin was more potent in inhibition of migration in the Matrigel tube formation assay (90%). Both compounds similarly inhibited proliferation (50-90%) of endothelial cells, and angiogenesis induced by vascular endothelial growth factor (80%) and glioma (55%) in the chorioallantoic membrane assay. These peptides were not toxic to endothelial cell cultures and caused no acute toxicity upon intravenous injection in mice, and were stable for 10-30 hours in human serum. The in vitro and in vivo potency of the peptides are consistent with conformational ensembles and "bioactive" space shared by obtustatin and viperistatin. These findings suggest that vimocin and vidapin can serve as dual a 1 b 1 /a 2 b 1 integrin antagonists in antiangiogenesis and cancer therapy.

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The Synthesis of Cystine Peptides by Iodine Oxidation of S‐Trityl‐cysteine and S‐Acetamidomethyl‐cysteine Peptides

Cited by 277 publications

References 45 publications

Post-translational Modifications of Endothelin Receptor B from Bovine Lungs Analyzed by Mass Spectrometry

Post-translational Modifications of Endothelin Receptor B from Bovine Lungs Analyzed by Mass Spectrometry

Optical spectroscopic elucidation of β‐turns in disulfide bridged cyclic tetrapeptides

Vimocin and Vidapin, Cyclic KTS Peptides, Are Dual Antagonists ofα₁β₁/α₂β₁Integrins with Antiangiogenic Activity

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The Synthesis of Cystine Peptides by Iodine Oxidation of S‐Trityl‐cysteine and S‐Acetamidomethyl‐cysteine Peptides

Cited by 277 publications

References 45 publications

Post-translational Modifications of Endothelin Receptor B from Bovine Lungs Analyzed by Mass Spectrometry

Post-translational Modifications of Endothelin Receptor B from Bovine Lungs Analyzed by Mass Spectrometry

Optical spectroscopic elucidation of β‐turns in disulfide bridged cyclic tetrapeptides

Vimocin and Vidapin, Cyclic KTS Peptides, Are Dual Antagonists ofα1β1/α2β1Integrins with Antiangiogenic Activity

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Vimocin and Vidapin, Cyclic KTS Peptides, Are Dual Antagonists ofα₁β₁/α₂β₁Integrins with Antiangiogenic Activity