Expression and characterization of the first kunitz domain of human tissue factor pathway inhibitor-2

Desheng Kong; Duan Ma; Hao Bai; Hongshen Guo; Xu Cai; Wei Mo; Qiqun Tang; Houyan Song

doi:10.1016/j.bbrc.2004.09.179

Expression and characterization of the first kunitz domain of human tissue factor pathway inhibitor-2

Desheng Kong, Duan Ma, Hao Bai, Hongshen Guo, Xu Cai, Wei Mo, Qiqun Tang, Houyan Song

School of Medicine

Research output: Contribution to journal › Article › peer-review

33 Scopus citations

Abstract

Human tissue factor pathway inhibitor-2 (hTFPI-2) has three kunitz domains whose structure and function are unclear. We expressed the first kunitz domain of hTFPI-2 (hTFPI-2/KD1) as functional form using Pichia pastoris and investigated its characterization. In the experiment, hTFPI-2/KD1 can inhibit the plasmin and trypsin activity and the Ki of hTFPI-2/KD1 towards plasmin (30 nM) and trypsin (50 nM) was determined as 10 and 7 nM by chromogenic assay, respectively. hTFPI-2/KD1 can also inhibit MMP-2 and MMP-9 in zymography assay. Furthermore, the inhibition of hTFPI-2/KD1 to the Matrigel invasion by HT-1080 is also described. This study provides a method to produce hTFPI-2/KD1 efficiently and some insights into the structure and function of hTFPI-2/KD1.

Original language	English (US)
Pages (from-to)	1179-1185
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	324
Issue number	4
DOIs	https://doi.org/10.1016/j.bbrc.2004.09.179
State	Published - Nov 26 2004

Keywords

Human tissue factor pathway inhibitor-2
Kunitz domain
Pichia pastoris
Plasmin
Protein expression
Purification
Trypsin

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.bbrc.2004.09.179

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title = "Expression and characterization of the first kunitz domain of human tissue factor pathway inhibitor-2",

abstract = "Human tissue factor pathway inhibitor-2 (hTFPI-2) has three kunitz domains whose structure and function are unclear. We expressed the first kunitz domain of hTFPI-2 (hTFPI-2/KD1) as functional form using Pichia pastoris and investigated its characterization. In the experiment, hTFPI-2/KD1 can inhibit the plasmin and trypsin activity and the Ki of hTFPI-2/KD1 towards plasmin (30 nM) and trypsin (50 nM) was determined as 10 and 7 nM by chromogenic assay, respectively. hTFPI-2/KD1 can also inhibit MMP-2 and MMP-9 in zymography assay. Furthermore, the inhibition of hTFPI-2/KD1 to the Matrigel invasion by HT-1080 is also described. This study provides a method to produce hTFPI-2/KD1 efficiently and some insights into the structure and function of hTFPI-2/KD1.",

keywords = "Human tissue factor pathway inhibitor-2, Kunitz domain, Pichia pastoris, Plasmin, Protein expression, Purification, Trypsin",

author = "Desheng Kong and Duan Ma and Hao Bai and Hongshen Guo and Xu Cai and Wei Mo and Qiqun Tang and Houyan Song",

note = "Funding Information: This study was supported in part by Shanghai Education Committee, Shanghai Scientific and Technology Committee, and 30271229 from The National Natural Sciences Foundation, China. Copyright: Copyright 2017 Elsevier B.V., All rights reserved.",

year = "2004",

month = nov,

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doi = "10.1016/j.bbrc.2004.09.179",

language = "English (US)",

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AU - Kong, Desheng

AU - Ma, Duan

AU - Bai, Hao

AU - Guo, Hongshen

AU - Cai, Xu

AU - Mo, Wei

AU - Tang, Qiqun

AU - Song, Houyan

N1 - Funding Information: This study was supported in part by Shanghai Education Committee, Shanghai Scientific and Technology Committee, and 30271229 from The National Natural Sciences Foundation, China. Copyright: Copyright 2017 Elsevier B.V., All rights reserved.

PY - 2004/11/26

Y1 - 2004/11/26

N2 - Human tissue factor pathway inhibitor-2 (hTFPI-2) has three kunitz domains whose structure and function are unclear. We expressed the first kunitz domain of hTFPI-2 (hTFPI-2/KD1) as functional form using Pichia pastoris and investigated its characterization. In the experiment, hTFPI-2/KD1 can inhibit the plasmin and trypsin activity and the Ki of hTFPI-2/KD1 towards plasmin (30 nM) and trypsin (50 nM) was determined as 10 and 7 nM by chromogenic assay, respectively. hTFPI-2/KD1 can also inhibit MMP-2 and MMP-9 in zymography assay. Furthermore, the inhibition of hTFPI-2/KD1 to the Matrigel invasion by HT-1080 is also described. This study provides a method to produce hTFPI-2/KD1 efficiently and some insights into the structure and function of hTFPI-2/KD1.

AB - Human tissue factor pathway inhibitor-2 (hTFPI-2) has three kunitz domains whose structure and function are unclear. We expressed the first kunitz domain of hTFPI-2 (hTFPI-2/KD1) as functional form using Pichia pastoris and investigated its characterization. In the experiment, hTFPI-2/KD1 can inhibit the plasmin and trypsin activity and the Ki of hTFPI-2/KD1 towards plasmin (30 nM) and trypsin (50 nM) was determined as 10 and 7 nM by chromogenic assay, respectively. hTFPI-2/KD1 can also inhibit MMP-2 and MMP-9 in zymography assay. Furthermore, the inhibition of hTFPI-2/KD1 to the Matrigel invasion by HT-1080 is also described. This study provides a method to produce hTFPI-2/KD1 efficiently and some insights into the structure and function of hTFPI-2/KD1.

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Expression and characterization of the first kunitz domain of human tissue factor pathway inhibitor-2

Abstract

Keywords

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