Fusion expression of DDR2 extracellular domain in insect cells and its purification and function characterization.

Fusion expression of DDR2 extracellular domain in insect cells and its purification and function characterization. Research Abstract Details 

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Fusion expression of DDR2 extracellular domain in insect cells and its purification and function characterization. Abstract Text:

Wei Zhang,Tianbing Ding,Jian Zhang,Jin Su,Jiangtian Yu,Jipeng Li,Fuyang Li,Chunmei Wang,Nannan Liu,Xinping Liu,Wenyu Ma,Libo Yao,

Discoidin domain receptor 2 (DDR2) is a kind of protein tyrosine kinases associated with cell proliferation and tumor metastasis, and collagen, a ligand for DDR2, up-regulates matrix metalloproteinase 1 (MMP-1) and MMP-2 expression in extracellular matrix (ECM). To investigate the role of DDR2 in cartilage destruction in rheumatoid arthritis (RA), we expressed the extracellular domain (ECD) of DDR2 (without signal peptide and transmembrane domain, designated DR) in insect cells, purified and characterized DR, hoping to use it as a specific antagonist of DDR2. By using Bac-To-Bac Expression System with a His tag, we successfully obtained the recombinant bacularvirus containing DDR2 ECD, purified it and characterized its function. The soluble fraction of DR was about 12% of the total fused protein. After chromatographic purification, DR with 92% purity was obtained. Competitive inhibition assay demonstrated that DR blocked the binding between DDR2 and natural DDR2 receptors on NIH3T3 and synovial cells. Results of RT-PCR, Western blotting, and gelatinase zymography showed that DR was capable of inhibiting MMP-1 and MMP-2 secretion from NIH3T3 and RA synoviocytes stimulated by collagen II. For MMP-1, inhibition was displayed at the levels of mRNA and protein, whereas for MMP-2 it was at the level of protein. These findings suggested that the expressed DR inhibited the activity of natural DDR2 and relevant MMP-1 and MMP-2 expression in RA synoviocytes and NIH3T3 cells provoked by collagen II.

Fusion expression of DDR2 extracellular domain in insect cells and its purification and function characterization. Publishing Authors By Initials

W Zhang,T Ding,J Zhang,J Su,J Yu,J Li,F Li,C Wang,N Liu,X Liu,W Ma,L Yao,

For similar animals: invertebrates: arthropods: insects: lepidoptera: moths: spodoptera research abstracts see: animals: invertebrates: arthropods: insects: lepidoptera: moths: spodoptera research

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Fusion expression of DDR2 extracellular domain in insect cells and its purification and function characterization. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of cellular biochemistry

VOLUME: 102

Page Numbers: 41-51

Journal Abbreviation: J. Cell. Biochem.

ISSN: 0730-2312

DAY: 1

MONTH: Sep

YEAR: 2007

Fusion expression of DDR2 extracellular domain in insect cells and its purification and function characterization. Information

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LANGUAGE: eng

NlmUniqueID: 8205768

Fusion expression of DDR2 extracellular domain in insect cells and its purification and function characterization. Keywords Mesh Terms:

KEYWORDS: Spodoptera

MESH TERMS: virology

Chemical & Substance for Abstract: Fusion expression of DDR2 extracellular domain in insect cells and its purification and function characterization. Information

Substance Name: Matrix Metalloproteinase 1

Registry Number: EC 3.4.24.7

Grant and Affiliation Information for Fusion expression of DDR2 extracellular domain in insect cells and its purification and function characterization.

AFFILIATION: Department of Microbiology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.

Country: United States

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MEDLINETA: J Cell Biochem

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