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

Research output: Contribution to journalArticlepeer-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 languageEnglish (US)
Pages (from-to)1179-1185
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume324
Issue number4
DOIs
StatePublished - Nov 26 2004
Externally publishedYes

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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