On the specific cleavage of cysteine containing peptides and proteins

Addition of cysteine to the double bonds of polydehydroalanine and copolymers of dehydroalanine (DHA) and methylcysteine, alanine, leucine or gamma- methyl-L-glutamate is accompanied by increased solubility and decreased molecular weight. This result is due to a peptide bond cleavage caused by formation of a thiazolidine as a consequence of nucleophilic attack by the sulfur atom on the preceding C=O group and subsequent splitting of the peptide bond by nucleophilic attack of an OH ion. This mechanism is predominant in alkaline media; in acid another mechanism is favoured; carbonyl-oxygen attacks the carbonyl-C-atom of the cysteine residue, forming a second ring system. Addition of one water molecule then yields two peptide fragments, one of them a terminal cysteine residue. Both mechanisms could be confirmed in the case of gamma-L-glutamyl-dehydroalanyl-glycine by adding cysteine. Furthermore, it could be shown that SH-glutathione is decomposed at alevated temperatures according to the two mechanisms mentioned. This SH-induced peptide bond cleavage can be used for selective peptide chain splitting of cysteine-containing polypeptides and proteins under relatively mild conditions.