Tissue Kallikreins and Related Enzymes: Characterization by Model Oligopeptides
The first purpose of this work was to obtain direct evidence that tissue kallikreins cleave arginyl bonds when the leaving group is Arg-Val, and on the contrary, do not split them when it is Arg-Pro; the second aim was to ascertain whether this specificity could be used as a criterion, for characterizing tissue kallikreins.
Two tetrapeptides A) Ac-Phe-Arg-Arg-Val-NH2 and B) Ac-Phe-Arg-Arg-Pro-NH2 were synthesized by the solid phase method and purified to homogeneity. They were used as substrates for homogeneous preparations of tissue and plasma kallikreins, as well as for some related serine proteases. Products identification and kinetic analyse were made by HPLC.
The hindering effect of the P2 Pro residue in the hydrolysis by tissue kallikreins was unequivocally demonstrated. Results showed also that enzymes which cleave the Arg-Arg bond in peptide A but do not hydrolyze peptide B, may be classified as tissue kallikreins.
KeywordsHigh Performance Liquid Chromatography High Performance Liquid Chromatography Proline Residue Tissue Kallikrein Solid Phase Method
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