Affinity labelling of proteinases with tryptic specificity by peptides with C-terminal lysine chloromethyl ketone |
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Authors: | John R Coggins William Kray and Elliott Shaw |
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Affiliation: | Biology Department, Brookhaven National Laboratory, Upton, N.Y. 11973, U.S.A. |
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Abstract: | Methods are described for the synthesis of peptides terminating in Lys-CH(2)Cl. The products were examined as affinity labels for several enzymes of trypsin-like specificity which are resistant to Tos-Lys-CH(2)Cl. In part, the inertness of the latter may be due to the sulphonamide group, since Z-Lys-CH(2)Cl was more effective. However, a number of tripeptides with C-terminal Lys-CH(2)Cl were superior in their ability to inactivate subtilisin, thrombin and plasma kallikrein. The possibility of developing enzyme-specific reagents selective for members within the trypsin-like group is demonstrated by Ala-Phe-Lys-CH(2)Cl, which readily inactivates plasma kallikrein but not thrombin. |
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