Expression, purification and structural characterization of functionally replete thrombospondin-1 type 1 repeats in a bacterial expression system.
Expression, purification and structural characterization of functionally replete thrombospondin-1 type 1 repeats in a bacterial expression system.
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DOI：
10.1016/j.pep.2011.07.010
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发表时间：
2011-12
影响因子：
1.6
通讯作者：
Silverstein, Roy L.
中科院分区：
文献类型：
--
作者：
Klenotic, Philip A.;Page, Richard C.;Misra, Saurav;Silverstein, Roy L.
关键词：
The matrix glycoprotein Thrombospondin-1 (TSP-1) is a prominent regulator of endothelial cells and angiogenesis. The anti-angiogenic and anti-tumorigenic properties of TSP-1 are in part mediated by the TSP-1 type 1 repeat domains 2 and 3, TSR(2,3). Here, we describe the expression and purification of human TSR(2,3) in milligram quantities from an Escherichia coli expression system. Microvascular endothelial cell migration assays and binding assays with a canonical TSP-1 ligand, histidine-rich glycoprotein (HRGP), indicate that recombinant TSR(2,3) exhibits anti-chemotactic and ligand binding properties similar to full length TSP-1. Furthermore, we determined the structure of E. coli expressed TSR(2,3) by X-ray crystallography at 2.4 Å and found the structure to be identical to the existing TSR(2,3) crystal structure determined from a Drosophila expression system. The TSR(2,3) expression and purification protocol developed in this study allows for facile expression of TSR(2,3) for biochemical and biophysical studies, and will aid in the elucidation of the molecular mechanisms of TSP-1 anti-angiogenic and anti-tumorigenic activities.
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DOI：
10.1083/jcb.138.3.707
发表时间：
1997-08-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Dawson DW;Pearce SF;Zhong R;Silverstein RL;Frazier WA;Bouck NP
通讯作者：
Bouck NP
影响因子：
15.9
作者：
Bornstein, P
通讯作者：
Bornstein, P
DOI：
10.1073/pnas.171460498
发表时间：
2001-10-23
影响因子：
11.1
作者：
Rodríguez-Manzaneque, JC;Lane, TF;Iruela-Arispe, ML
通讯作者：
Iruela-Arispe, ML
影响因子：
3.4
作者：
Lu, Chao;Kerbel, Robert S.
通讯作者：
Kerbel, Robert S.
DOI：
10.1111/j.1432-1033.1981.tb05655.x
发表时间：
1981-01-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
RYLATT, DB;SIA, DY;PARISH, CR
通讯作者：
PARISH, CR