Expression of the cell‐binding domain of human fibronectin in E. coli

FEBS Letters

Thiery, Jean Paul²  Kang, Mohinder S.¹  Obara, Masanobu¹  Yamada, Kenneth M.¹  Rocher-Dufour, Sylvie²  Kornblihtt, Alberto³ 

[1] Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA;Institut d'Embryologie, CNRS et College de France, 94130 Nogent sur Marne Cedex, France;Instituto de Investigaciones en Ingenieria Genetica y Biologia Molecular (INGEBI-CONICET), Buenos Aires, Argentina

关键词: Fibronectin;    cDNA;    Cell adhesion;    (E. coli. Human);   

DOI  :  10.1016/0014-5793(87)81502-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Two cDNA subfragments containing the cell-attachment site of human fibronectin (FN) were expressed as β-galactosidase fusion proteins in E. coli. The products were purified to homogeneity by monoclonal antibody affinity chromatography and assayed for activity in a standard cell-adhesion assay. A fusion protein containing an 80 kDa fragment of human FN appeared functionally equivalent to intact FN purified from human plasma, whereas a truncated fusion protein of 33 kDa still containing a previously postulated cell-attachment site was approx. 50-fold less active. Our study establishes a system for analyzing adhesive protein function by DNA manipulation, rules out any major role for eukaryotic post-translational modifications in FN adhesive function, and localizes additional functional activity to a 1.3 kb region.

【 授权许可】

Unknown