Characterization of a cDNA for Rhesus Monkey Protein C Inhibitor – Evidence for N-Terminal Involvement in Heparin Stimulation

Klaus-P Radtke; José A Fernández; Bruno O Villoutreix; Judith S Greengard; John H Griffin

doi:10.1055/s-0038-1649885

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1995; 74(04): 1079-1087
DOI: 10.1055/s-0038-1649885

Original Article

Coagulation

Schattauer GmbH Stuttgart

Characterization of a cDNA for Rhesus Monkey Protein C Inhibitor – Evidence for N-Terminal Involvement in Heparin Stimulation

Klaus-P Radtke

Department of Molecular and Experimental Medicine, Department of Vascular Biology, The Scrlpps Research Institute, La Jolla, California, CA, USA

,

José A Fernández

Department of Molecular and Experimental Medicine, Department of Vascular Biology, The Scrlpps Research Institute, La Jolla, California, CA, USA

,

Bruno O Villoutreix

Department of Molecular and Experimental Medicine, Department of Vascular Biology, The Scrlpps Research Institute, La Jolla, California, CA, USA

,

Judith S Greengard

Department of Molecular and Experimental Medicine, Department of Vascular Biology, The Scrlpps Research Institute, La Jolla, California, CA, USA

,

John H Griffin

Department of Molecular and Experimental Medicine, Department of Vascular Biology, The Scrlpps Research Institute, La Jolla, California, CA, USA

› Institutsangaben

Abstract

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Summary

cDNAs for protein C inhibitor (PCI) were cloned from human and rhesus monkey[ ¹ ] liver RNAs by reverse transcription and polymerase chain reaction (PCR) amplification. Sequencing showed that rhesus monkey and human PCI cDNAs were 93% identical. Predicted amino acid sequences differed at 26 of 387 residues. Pour of these differences (T352M, N359S, R362K, L3631) were in the reactive center loop that is important for inhibitory specificity, and two were in the N-terminal helix (M8T, E13K) that is implicated in glycosaminoglycan binding. PCI in human or rhesus monkey plasma showed comparable inhibitory activity towards human activated protein C in the presence of 10 U/ml heparin. However, maximal acceleration of the inhibition of activated protein C required 5-fold lower heparin concentration for rhesus monkey than for human plasma, consistent with the interpretation that the additional positive charge (E13K) in a putative-heparin binding region increased the affinity for heparin.

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