EXPRESSION IN ESCHERICHIA-COLI, PHOSPHORYLATION WITH CAMP-DEPENDENT PROTEIN-KINASE AND PROTEOLYSIS BY CALPAIN OF A 71-KDA DOMAIN OF HUMAN ENDOTHELIAL ACTIN-BINDING PROTEIN

A middle region of human endothelial actin-binding protein (ABP) was s ubcloned and expressed in the pT7-7/E. coli BL21 (DE3) system. As pred icted by the amino acid sequence this 71 kD truncated protein (residue s 1717-2360) contained a calpain cleavage site and two of the three pr esumptive cAMP-dependent protein kinase phosphorylation sites. This pe ptide fragment comprised all the elements needed to confer stability a gainst calpain proteolysis to ABP after PKA phosphorylation. (C) 1997 Academic Press.