PMID: 6967737Aug 7, 1980Paper

Purification of guinea-pig plasma prekallikrein. Activation by prekallikrein activator derived from guinea-pig skin

Biochimica Et Biophysica Acta
T YamamotoT Kambara

Abstract

Prekallikrein was purified from guinea-pig plasma. The prekallikrein appeared homogeneous as a single-chain protein on polyacrylamide gels in the presence of sodium dodecyl sulfate (SDS) and beta-mercaptoethanol. The apparent molecular weight was 82 000 by SDS-polyacrylamide gel electrophoresis, 99 000 by gel filtration on a Sephadex G-150 column and 84 500 (protein part) by amino acid analysis. The isoelectric point was approx. 9.0. The purification method yielded 3.8 mg (A280 3.800) of prekallikrein from 500 ml of plasma. Kallikrein was generated from the prekallikrein by limited proteolytic action of a prekallikrein activator which was derived from guinea-pig skin. From analysis using SDS-polyacrylamide gel electrophoresis, the kallikrein has two fragments with apparent molecular weights of 52 000 and 40 000 which are linked by disulfide bond(s). The 40 000 molecular weight fragment was shown to incorporate [3H]diisopropylfluorophosphate. The kallikrein hydrolyzed the synthetic substrates containing the Phe-Arg sequence at the COOH-terminal, and it cleaved carbobenzyloxy-Phe-Arg-4-methylcoumaryl-7-amide more readily than Pro-Phe-Arg-methylcoumaryl-7-amide. The Km for the kallikrein with carbobenzyloxy-Phe-Arg-methylcoumaryl ...Continue Reading

References

Jan 1, 1979·Seminars in Thrombosis and Hemostasis·J H Griffin, C G Cochrane
Mar 1, 1978·Thrombosis Research·M J GallimoreH Stormorken
Jan 1, 1976·Methods in Enzymology·R W Colman, A Bagdasarian
Jan 1, 1972·The Journal of Experimental Medicine·K D Wuepper, C G Cochrane
Jan 1, 1972·Methods in Enzymology·K WeberM Osborn
Apr 29, 1969·Biochemical and Biophysical Research Communications·H Matsubara, R M Sasaki

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Citations

Aug 30, 1991·Clinica Chimica Acta; International Journal of Clinical Chemistry·Y ShibuyaH Okabe
Jan 1, 1984·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·J C Wriston
Dec 30, 1999·Immunopharmacology·Y ShibuyaT Yamamoto
Jun 1, 1991·Japanese Journal of Cancer Research : Gann·Y MatsumuraH Maeda
Aug 1, 1984·Preparative Biochemistry·G S Bedi, N Back
Jul 26, 1991·Biochimica Et Biophysica Acta·Y ShibuyaT Yamamoto
Aug 11, 1988·Biochimica Et Biophysica Acta·T YamamotoC G Cochrane
Jun 11, 1993·Biochimica Et Biophysica Acta·M M KhanT Kambara
Feb 1, 1992·Biochimica Et Biophysica Acta·T Imamura, T Kambara

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