Determination of the N-terminal amino acid sequence of the purified prothrombin from a patient with liver cirrhosis.
The reasons for the decreased functional activity of prothrombin in liver diseases are still speculative. When a highly purified preparation of prothrombin from a patient with liver cirrhosis is available, the cause of prothrombin abnormalities may be researched on a molecular basis. In this study, prothrombin (6.7 mg) was purified from the ascites fluid (1130 mL) of a patient with liver cirrhosis by barium citrate adsorption, ammonium sulfate elution, DEAE Sephacel and Heparin Sepharose CL-6B column chromatography steps. The molecular weight of this prothrombin was the same as that of normal prothrombin purified from a normal plasma pool. The specific activities were found to be 3.36 U/mg in the one stage clotting assay and 28.9 U/mg in the staphylocoagulase/chromogenic substrate assay, while the normal prothrombin specific activities were 3.92 U/mg and 30.1 U/mg respectively. When N-terminal amino acid sequence analysis was carried out, it was seen that the first 20 residues were identical to the normal human prothrombin excepting the Gla at position #14.
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Department of Biochemistry, University of Marmara Faculty of Pharmacy, Istanbul, Turkey. ariza@anet.net.tr
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Yardimci
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Sardana
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D015055:1-Carboxyglutamic Acid / Q000032:analysis*; D000595:Amino Acid Sequence; D001202:Ascitic Fluid / Q000737:chemistry*; D018698:Glutamic Acid / Q000032:analysis; D006474:Hemorrhagic Disorders / Q000209:etiology; D006801:Humans; D008103:Liver Cirrhosis / Q000150:complications / Q000378:metabolism*; D008970:Molecular Weight; D011499:Protein Processing, Post-Translational; D011516:Prothrombin / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011517:Prothrombin Time; D014812:Vitamin K / Q000378:metabolism
D014812:Vitamin K; D018698:Glutamic Acid; D015055:1-Carboxyglutamic Acid; D011516:Prothrombin