Proteolytic cleavage of the puromycin-sensitive aminopeptidase generates a substrate binding domain.

摘要: Abstract The puromycin-sensitive aminopeptidase was found to be resistant proteolysis by trypsin, chymotrypsin, and protease V8 but cleaved into an N-terminal 60-kDa fragment a C-terminal 33-kDa proteinase K. two K fragments remain associated retained enzymatic activity. Attempts express the in Escherichia coli produced inclusion bodies. A hexa-histidine fusion protein of solubilized from bodies with urea refolded removal through dialysis. devoid activity as assayed arginine–β-naphthylamide. However, bound substrate dynorphin A(1-9) stoichiometry 0.5 mol/mol K0.5 value 50 μM. Dynorphin binding competitively inhibited B(1-9), not des-Tyr1-leucine-enkephalin, poor for enzyme.