Molecular cloning, expression, and characterization of bovine tissue factor pathway inhibitor-2

被引:5
作者
Du, X
Deng, FM
Chand, HS
Kisiel, W [1 ]
机构
[1] Univ New Mexico, Hlth Sci Ctr, Dept Pathol, Albuquerque, NM 87131 USA
[2] NYU, Sch Med, Ronald Perelman Dept Dermatol, Epithelial Biol Unit, New York, NY 10016 USA
关键词
tissue factor pathway inhibitor-2; cDNA sequence; serine proteinase inhibitor;
D O I
10.1016/S0003-9861(03)00332-1
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
Human tissue factor pathway inhibitor-2 (TFPI-2) is a matrix-associated Kunitz-type serine proteinase inhibitor that is secreted by all cells of the vasculature, and presumably plays a role in the regulation of plasmin-mediated matrix remodeling. In this report, we describe the cloning and expression of a full-length cDNA for bovine TFPI-2 that exhibits 72% sequence identity with that of human TFPI-2. Following a 22 residue signal peptide, the mature protein contains 212 amino acids with 18 cysteines, three putative N-glycosylation sites, and one putative O-glycosylation site. The deduced sequence of mature bovine TFPI-2 revealed a short acidic amino-terminal region, three tandem Kunitz-type domains, and a carboxy-terminal tail highly enriched in basic amino acids. Recombinant bovine TFPI-2 was expressed in HEK 293 cells and resolved into two isoforms, designated as alpha-TFPI-2 (M-r 33 kDa) and beta-TFPI-2 (M-r 31 kDa), which presumably represent differentially glycosylated forms of the inhibitor. Similar to human TFPI-2, both bovine TFPI-2 isoforms exhibited strong inhibitory activity towards trypsin and plasmin, and weak inhibitory activity towards the factor VIIa-tissue factor complex. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码:96 / 104
页数:9
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