Expression and characterization of Kunitz domain 3 and C-terminal of human tissue factor pathway inhibitor-2

被引：6

作者：

Zhu, Lina ^{[1
]}

Wang, Jiping ^{[2
]}

Mu, Jingui ^{[2
]}

Wang, Huijun ^{[3
]}

Zhang, Chenqi ^{[1
]}

Wang, Jue ^{[1
]}

Liu, Xingang ^{[1
]}

Yan, Xiaomin ^{[1
]}

Dai, Linsen ^{[1
]}

Ma, Duan ^{[2
,3
]}

机构：

[1] Fudan Univ, Ctr Anal & Measurement, Shanghai 200433, Peoples R China

[2] Fudan Univ, Shanghai Med Coll, Minist Educ, Key Lab Mol Med, Shanghai 200032, Peoples R China

[3] Fudan Univ, Inst Biomed Sci, Shanghai 200032, Peoples R China

来源：

ACTA BIOCHIMICA ET BIOPHYSICA SINICA | 2009年 / 41卷 / 11期

基金：

中国国家自然科学基金;

关键词：

Kunitz domain 3 and C-terminal of hTFPI-2; heparin; secondary structure; Pichia pastoris; SERINE PROTEINASE-INHIBITORS; YEAST PICHIA-PASTORIS; PROTEASE INHIBITORS; HEPARIN-BINDING; CDNA CLONING; CELLS; RECOMBINANT; MECHANISM; SPECTRA; GENE;

D O I：

10.1093/abbs/gmp089

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 (hTFPI-2) is a serine protease inhibitor and its inhibitory activity is enhanced by heparin. The Kunitz domain 3 and C-terminal of hTFPI-2 (hTFPI-2/KD3C), which has the activity toward heparin calcium, have been successfully expressed in Pichia pastoris and purified by SP-Sepharose and heparin-Sepharose chromatography. The Fourier transformed infrared spectroscopy (FTIR), Raman spectroscopy, and circular dichroism (CD) experiment results implied that hTFPI-2/KD3C contained small contents of alpha-helix and beta-strand, but large amounts of random coil and two kinds of disulfide bonds, gauche-gauche-gauche (ggg) and trans-gauche-trans (tgt). The interaction of hTFPI-2/KD3C with heparin calcium was investigated by CD. It was found that heparin calcium induced beta-strands in hTFPI-2/KD3C to different extents depending on the ratio of hTFPI-2/KD3C and heparin calcium.

引用

页码：948 / 954

页数：7

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