Preparation and characterization of a novel chimeric protein VEGI-CTT in Escherichia coli

被引:7
作者
Cai, Jiping [1 ]
Wei, Ruili [1 ]
Cheng, Jinwei [1 ]
机构
[1] Second Mil Med Univ, Affiliated Hosp 2, Dept Ophthalmol, Shanghai 200433, Peoples R China
来源
JOURNAL OF BIOMEDICINE AND BIOTECHNOLOGY | 2008年
关键词
D O I
10.1155/2008/564969
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Vascular endothelial cell growth inhibitor ( VEGI) is a recently identified antiangiogenic cytokine that belongs to the TNF superfamily, and could effectively inhibit endothelial cell proliferation and angiogenesis. Synthetic peptide CTT (CTTHWGFTLC) has been found to suppress invasion and migration of both tumor and endothelial cells by potent and selective inhibition of MMP-2 and MMP-9. To prepare chimeric protein VEGI-CTT for more potent antitumor therapy, the recombinant expression vector pET-VEGI-CTT was constructed. This fusion protein was expressed in inclusion bodies in E. coli BL21 (DE3), and was refolded and purified by immobilized metal affinity chromatography using His-tag. Purified VEGI-CTT protein was characterized by proliferation assays of the endothelial cells and casein degradation assay in vitro. The results demonstrated that chimeric protein VEGI-CTT had a potent activity of antiangiogenesis through inhibiting the proliferation of endothelial cells, and could effectively reduce the activity of MMP-2 and MMP-9. The preliminarily in vivo study demonstrated that chimeric protein VEGI-CTT had more potent antitumor activity than VEGI and/or CTT peptide against CA46 human lymphoma xenografts in nude mice. Thus, these facts that are derived from the present study suggest that the chimeric protein VEGI-CTT may be used for tumor therapy in the future. Copyright (c) 2008 Jiping Cai et al.
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页数:9
相关论文
共 30 条
  • [1] Aggarwal BB, 1996, EUR CYTOKINE NETW, V7, P93
  • [2] REMOVAL OF SODIUM DODECYL-SULFATE FROM PROTEINS AND PEPTIDES BY GEL-FILTRATION
    AMONS, R
    SCHRIER, PI
    [J]. ANALYTICAL BIOCHEMISTRY, 1981, 116 (02) : 439 - 443
  • [3] A novel secreted splice variant of vascular endothelial cell growth inhibitor
    Chew, LJ
    Pan, HG
    Yu, JY
    Tian, S
    Huang, WQ
    Zhang, JY
    Pang, S
    Li, LY
    [J]. FASEB JOURNAL, 2002, 16 (03) : 742 - +
  • [4] Clark EDB, 1998, CURR OPIN BIOTECH, V9, P157
  • [5] Discovery and development of bevacizumab, an anti-VEGF antibody for treating cancer
    Ferrara, N
    Hillan, KJ
    Gerber, HP
    Novotny, W
    [J]. NATURE REVIEWS DRUG DISCOVERY, 2004, 3 (05) : 391 - 400
  • [6] Angiogenic zip code
    Folkman, J
    [J]. NATURE BIOTECHNOLOGY, 1999, 17 (08) : 749 - 749
  • [7] Endogenous angiogenesis inhibitors
    Folkman, J
    [J]. APMIS, 2004, 112 (7-8) : 496 - 507
  • [8] Construction and high-level expression of a single-chain Fv antibody fragment specific for acidic isoferritin in Escherichia coli
    Guo, JQ
    You, SY
    Li, L
    Zhang, YZ
    Huang, JN
    Zhang, CY
    [J]. JOURNAL OF BIOTECHNOLOGY, 2003, 102 (02) : 177 - 189
  • [9] A His-tag based immobilization method for the preparation and reconstitution of apoflavoproteins
    Hefti, MH
    Milder, FJ
    Boeren, S
    Vervoort, J
    van Berkel, WJH
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 2003, 1619 (02): : 139 - 143
  • [10] [侯卫红 Hou Weihong], 2004, [郑州大学学报. 医学版, Journal of Zhengzhou University.Medical Sciences], V39, P51