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
收藏
页码：948 / 954
页数：7
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