Oxidative folding of synthetic polypeptidesS-protected astert-butylthio derivatives
| Autor: | Antonio Silvio Verdini, Mario Roggero, Silvia Terenzi, Vincent Brossard, Giampietro Corradin |
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| Rok vydání: | 2008 |
| Předmět: |
Pharmacology
chemistry.chemical_classification Stereochemistry Oxidative folding Organic Chemistry Peptide General Medicine Cleavage (embryo) Biochemistry Folding (chemistry) chemistry.chemical_compound Chaotropic agent Solid-phase synthesis chemistry Structural Biology Drug Discovery Peptide synthesis Molecular Medicine Organic chemistry Molecular Biology Cysteine |
| Zdroj: | Journal of Peptide Science. 14:1271-1282 |
| ISSN: | 1075-2617 |
| Popis: | A new method for oxidative folding of synthetic polypeptides assembled by stepwise solid phase synthesis is introduced. Folding is obtained in excellent yields by reacting S-tert-butylthiolated polypeptides with a 100-fold molar excess of cysteine at 37 °C in a slightly alkaline buffer containing chaotropic salts, and in the presence of air-oxygen. This novel protocol has been applied to the folding of S-tert-butylthiolated human thymus and activation-regulated chemokine (hu-TARC) derivatives as well as to larger segments of Plasmodium falciparum and Plasmodium berghei circumsporozoite proteins. Folded P. falciparum polypeptides have been used as substrates of endoproteinase Glu-C (Glu-C) and endoproteinase Asp-N (Asp-N) in an attempt to identify their disulfide connectivities. Particular practical advantages of the present method are (i) easy purification and storage of the S-protected peptide derivatives, (ii) elimination of the risk of cysteine alkylation during the acidolytic cleavage deprotection and resin cleavage steps, (iii) possibility to precisely evaluate the extent of folding and disulfide bond formation by mass spectrometry, and (iv) facile recovery of the final folded product. Copyright © 2008 European Peptide Society and John Wiley & Sons, Ltd. |
| Databáze: | OpenAIRE |
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