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 (30nM) and trypsin (50nM) was determined as 10 and 7nM 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.
authors.affiliation: Key Laboratory of Molecular Medicine, Ministry of Education, Dong'an Road 130#, Fudan University, Shanghai 200032, China.
authors.forename: Desheng
authors.initials: D
authors.lastname: Kong
authors.forename: Duan
authors.initials: D
authors.lastname: Ma
authors.forename: Hao
authors.initials: H
authors.lastname: Bai
authors.forename: Hongshen
authors.initials: H
authors.lastname: Guo
authors.forename: Xu
authors.initials: X
authors.lastname: Cai
authors.forename: Wei
authors.initials: W
authors.lastname: Mo
authors.forename: Qiqun
authors.initials: Q
authors.lastname: Tang
authors.forename: Houyan
authors.initials: H
authors.lastname: Song
mesh_terms: D045744:Cell Line, Tumor; D005341:Fibrinolysin / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020782:Matrix Metalloproteinases / Q000378:metabolism; D009361:Neoplasm Invasiveness; D010843:Pichia / Q000235:genetics; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000032:analysis / Q000302:isolation & purification / Q000494:pharmacology; D014357:Trypsin / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D011994:Recombinant Proteins; C021240:tissue-factor-pathway inhibitor 2; D014357:Trypsin; D005341:Fibrinolysin; D020782:Matrix Metalloproteinases