PURIFICATION AND CHARACTERIZATION OF 2 FORMS OF SOLUBLE THROMBOMODULIN FROM HUMAN URINE

被引：26

作者：

JACKSON, DE

TETAZ, TJ

SALEM, HH

MITCHELL, CA

机构：

[1] BOX HILL HOSP,MONASH MED SCH,DEPT MED,MELBOURNE 3128,AUSTRALIA

[2] BAKER MED RES INST,MELBOURNE,VIC,AUSTRALIA

来源：

EUROPEAN JOURNAL OF BIOCHEMISTRY | 1994年 / 221卷 / 03期

关键词：

D O I：

10.1111/j.1432-1033.1994.tb18827.x

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We have isolated and characterized two forms of soluble thrombomodulin from human urine. The purification procedure consisted of ultrafiltration, chromatography on DEAE-Sepharose, affinity chromatography on diisopropyl-phosphate - thrombin and/or monoclonal anti-thrombomodulin IgG affigel followed by reverse-phase HPLC. An active soluble form of thrombomodulin was purified 1600-fold from 34-1 urine. The purified protein migrated as a doublet, with molecular mass 76/72 kDa under reducing conditions and 63/57 kDa under non-reducing conditions as determined by SDS/PAGE. Amino acid analysis of the 63/57-kDa soluble thrombomodulin confirmed sequence identity with human thrombomodulin and demonstrated N-terminal heterogeneity. Compared to membrane-type thrombomodulin, the purified 63/57-kDa soluble thrombomodulin was more active as a cofactor for protein-C activation. The second major thrombomodulin fragment in urine is an inactive 35-kDa thrombomodulin polypeptide derived from the N-terminal extracellular region of thrombomodulin.

引用

页码：1079 / 1087

页数：9

共 28 条

[1]

BRISSON C, 1992, THROMB HAEMOSTASIS, V68, P737

[2]

BURNETTE WN, 1981, ANAL BIOCHEM, V112, P195, DOI 10.1016/0003-2697(81)90281-5

[3] IDENTIFICATION OF AN ENDOTHELIAL-CELL COFACTOR FOR THROMBIN-CATALYZED ACTIVATION OF PROTEIN-C [J].