Resistance to inhibition by alpha-1-anti-trypsin and species specificity of a chimeric human/bovine protein C.
Human activated protein C (APC) has been shown to be physiologically susceptible to inhibition by the abundant serpin inhibitor alpha-1-anti-trypsin (AAT). Studies on the inactivation by AAT [Heeb, M. J., & Griffin, J. H. (1988) J. Biol. Chem. 263, 11613-11616] have shown that the calculated rate of this inactivation matches that of the observed half-life of APC in vivo [Wydro, R., Oppenheimer, C., Rodger, R., & Miemi, S. (1988) Clin. Res. 36, 329A] and complex formation therefore probably represents a physiologic regulation process for APC. In this study we observed that bovine APC, in contrast to human APC, is nearly completely resistant to inactivation by human AAT. An additional difference between human and bovine APC is that human APC is a potent anticoagulant in human plasma, whereas bovine APC is only minimally active in human plasma. These functional differences exist despite considerable structural similarity between the human and bovine molecules. In order to identify specific molecular regions responsible for function, a chimeric molecule consisting of the light chain of human protein C (PC) and the heavy chain of bovine PC was constructed, expressed, and characterized. The activated chimeric PC is similar to human APC in having potent anticoagulant activity in human plasma, but displays nearly identical resistance to AAT inhibition with the bovine molecule. The similarity between the chimeric and bovine molecules in resistance to AAT inhibition indicates that the structural determinants for inhibitor interactions reside within the heavy chain (serine protease) domain.(ABSTRACT TRUNCATED AT 250 WORDS)
Holly
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ZymoGenetics, Inc., Seattle, Washington 98105.
Foster
D C
DC
D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D002417:Cattle; D002460:Cell Line; D004789:Enzyme Activation / Q000187:drug effects; D006207:Half-Life; D006801:Humans; D011486:Protein C / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors; D014162:Transfection; D000515:alpha 1-Antitrypsin / Q000494:pharmacology*
D011486:Protein C; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D000515:alpha 1-Antitrypsin