Expression and characterization of Kunitz domain 3 and C-terminal of human tissue factor pathway inhibitor-2
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 p...
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| Published in | Acta biochimica et biophysica Sinica Vol. 41; no. 11; pp. 948 - 954 |
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| Main Authors | , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
China
Oxford University Press
01.11.2009
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1672-9145 1745-7270 1745-7270 |
| DOI | 10.1093/abbs/gmp089 |
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| Abstract | 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 α-helix and β-strand, but large amounts of random coil and two kinds of disulfide bonds, gauche-gauche-gauche （ggg） and trans-gauchetrans （tgt）. The interaction of hTFPI-2/KD3C with heparin calcium was investigated by CD. It was found that heparin calcium induced β-strands in hTFPI-2/ KD3C to different extents depending on the ratio of hTFPI-2/KD3C and heparin calcium. |
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| AbstractList | 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 α-helix and β-strand, but large amounts of random coil and two kinds of disulfide bonds, gauche-gauche-gauche （ggg） and trans-gauchetrans （tgt）. The interaction of hTFPI-2/KD3C with heparin calcium was investigated by CD. It was found that heparin calcium induced β-strands in hTFPI-2/ KD3C to different extents depending on the ratio of hTFPI-2/KD3C and heparin calcium. 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 Cterminal of hTFPI-2 (hTFPI-2/KD3C), which has the activity toward heparin calcium, have been successfully expressed in Pichia pastoris and purified by SPSepharose 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-gauchetrans (tgt). The interaction of hTFPI-2/KD3C with heparin calcium was investigated by CD. It was found that heparin calcium induced b-strands in hTFPI-2/ KD3C to different extents depending on the ratio of hTFPI-2/KD3C and heparin calcium. 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 -helix and -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 -strands in hTFPI-2/KD3C to different extents depending on the ratio of hTFPI-2/KD3C and heparin calcium. 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 Cterminal of hTFPI-2 (hTFPI-2/KD3C), which has the activity toward heparin calcium, have been successfully expressed in Pichia pastoris and purified by SPSepharose 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-gauchetrans (tgt). The interaction of hTFPI-2/KD3C with heparin calcium was investigated by CD. It was found that heparin calcium induced b-strands in hTFPI-2/ KD3C to different extents depending on the ratio of hTFPI-2/KD3C and heparin calcium.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 Cterminal of hTFPI-2 (hTFPI-2/KD3C), which has the activity toward heparin calcium, have been successfully expressed in Pichia pastoris and purified by SPSepharose 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-gauchetrans (tgt). The interaction of hTFPI-2/KD3C with heparin calcium was investigated by CD. It was found that heparin calcium induced b-strands in hTFPI-2/ KD3C to different extents depending on the ratio of hTFPI-2/KD3C and heparin calcium. |
| Author | Lina Zhu Jiping Wang Jingui Mu Huijun Wang Chenqi Zhang Jue Wang Xingang Liu Xiaomin Yan Linsen Dai Duan Ma |
| AuthorAffiliation | Center of Analysis and Measurement, Fudan University, Shanghai 200433, China Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai Medical College, Fudan University, Shanghai 200032, China Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China |
| Author_xml | – sequence: 1 givenname: Lina surname: Zhu fullname: Zhu, Lina organization: 1 Center of Analysis and Measurement, Fudan University, Shanghai 200433, China – sequence: 2 givenname: Jiping surname: Wang fullname: Wang, Jiping organization: 2 Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai Medical College, Fudan University, Shanghai 200032, China – sequence: 3 givenname: Jingui surname: Mu fullname: Mu, Jingui email: duanma@shmu.edu.cn organization: 2 Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai Medical College, Fudan University, Shanghai 200032, China – sequence: 4 givenname: Huijun surname: Wang fullname: Wang, Huijun organization: 3 Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China – sequence: 5 givenname: Chenqi surname: Zhang fullname: Zhang, Chenqi organization: 1 Center of Analysis and Measurement, Fudan University, Shanghai 200433, China – sequence: 6 givenname: Jue surname: Wang fullname: Wang, Jue organization: 1 Center of Analysis and Measurement, Fudan University, Shanghai 200433, China – sequence: 7 givenname: Xingang surname: Liu fullname: Liu, Xingang organization: 1 Center of Analysis and Measurement, Fudan University, Shanghai 200433, China – sequence: 8 givenname: Xiaomin surname: Yan fullname: Yan, Xiaomin organization: 1 Center of Analysis and Measurement, Fudan University, Shanghai 200433, China – sequence: 9 givenname: Linsen surname: Dai fullname: Dai, Linsen email: lsdai@fudan.edu.cn organization: 1 Center of Analysis and Measurement, Fudan University, Shanghai 200433, China – sequence: 10 givenname: Duan surname: Ma fullname: Ma, Duan email: duanma@shmu.edu.cn organization: 2 Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai Medical College, Fudan University, Shanghai 200032, China |
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| CitedBy_id | crossref_primary_10_1016_j_pep_2016_03_003 crossref_primary_10_1002_jgm_3166 crossref_primary_10_1016_j_thromres_2010_02_017 crossref_primary_10_1177_1076029611429785 crossref_primary_10_1186_s13071_015_1022_z crossref_primary_10_1371_journal_pntd_0009007 |
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| Snippet | 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-... 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... |
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| SubjectTerms | Glycoproteins - chemistry Glycoproteins - isolation & purification Glycoproteins - metabolism Heparin - chemistry Humans Pichia - genetics Pichia - metabolism Protein Binding Protein Engineering - methods Protein Structure, Tertiary Recombinant Proteins - chemistry Recombinant Proteins - metabolism 丝氨酸蛋白酶抑制剂 傅立叶变换红外光谱 抑制物 组织因子途径 肝素钙 |
| Title | Expression and characterization of Kunitz domain 3 and C-terminal of human tissue factor pathway inhibitor-2 |
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