Detection of heparin-like glycosaminoglycans in normal human plasma by polyacrylamide-gel electrophoresis

被引:9
作者
Cavari, S [1 ]
Vannucchi, S [1 ]
机构
[1] UNIV FLORENCE,INST GEN PATHOL,I-50134 FLORENCE,ITALY
关键词
plasma; heparin; antithrombin III; electrophoresis;
D O I
10.1016/0009-8981(96)06331-0
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
This paper describes a procedure that allows detection of endogenous heparin present in normal human plasma by polyacrylamide-gel electrophoresis (PAGE). Plasma was submitted to proteolysis: an antithrombin III-dependent and heparinase I-sensitive anticoagulant activity was demonstrated in the supernatant of the digest. The supernatant was submitted to sequential fractionation with increasing concentrations of ethanol (25%, 50%, 60% and 65%, by vol.). Fractions were analyzed by PAGE for both glycosaminoglycan (GAG) and protein content. GAGs were detected by gradient PAGE (24-30%). The fraction obtained by 60% ethanol precipitation contained heparinase I-sensitive GAG. We show that GAGs co-precipitate with proteins. The SDS-PAGE of the material resulting from proteolytic digestion and subsequent ethanol fractionation, revealed three major bands. These peptides co-precipitated with plasma GAGs, mainly with the fraction obtained by 60% ethanol. We discuss the possibility that circulating endogenous heparin interacts with such peptides.
引用
收藏
页码:159 / 170
页数:12
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