Human Placental Anticoagulant Protein: Isolation and Characterization

Takayuki Funakoshi, Ronald L. Heimark, Lee E. Hendrickson, Brad A. McMullen, Kazuo Fujikawa

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202 Scopus citations

Abstract

An anticoagulant protein was purified from the soluble fraction of human placenta by ammonium sulfate precipitation and column chromatography on DEAE-Sepharose, Sephadex G-75, and Mono S (Pharmacia). The yield of the purified protein was approximately 20 mg from one placenta. The purified protein gave a single band by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a moelcular weight of 36 500. This protein prolonged the clotting time of normal plasma when clotting was induced either by brain thromboplastin or by kaolin in the presence of cephalin and Ca2+. It also prolonged the factor Xa induced clotting time of platelet-rich plasma but did not affect thrombin-induced conversion of fibrinogen to fibrin. The purified placental protein completely inhibited the prothrombin activation by reconstituted prothrombinase, a complex of factor Xa-factor Va-phospholipid-Ca2+. The placenta inhibitor had no effect on prothrombin activation when phospholipid was omitted from the above reaction. Also, it neither inhibited the amidolytic activity of factor Xa, nor did it bind to factor Xa. The placenta inhibitor, however, did bind specifically to phospholipid vesicles (20% phosphatidylserine and 80% phosphatidylcholine) in the presence of calcium ions. These results indicate that the placental anticoagulant protein (PAP) inhibits coagulation by binding to phospholipid vesicles. The amino acid sequences of three cyanogen bromide fragments of PAP aligned with those of two distinct regions of lipocortin I and II with a high degree of homology, showing that PAP is a member of the lipocortin family.

Original languageEnglish (US)
Pages (from-to)5572-5578
Number of pages7
JournalBiochemistry
Volume26
Issue number17
DOIs
StatePublished - Jan 1 1987
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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    Funakoshi, T., Heimark, R. L., Hendrickson, L. E., McMullen, B. A., & Fujikawa, K. (1987). Human Placental Anticoagulant Protein: Isolation and Characterization. Biochemistry, 26(17), 5572-5578. https://doi.org/10.1021/bi00391a053