Parole chiave: Recombinant Protein, Vasculotropin A, Vasculotropin Receptor 1, Vegfa Protein, Human, Article, Binding Site, Biosynthesis, Cell Culture, Chemistry, Drug Development, Endothelium Cell, Genetics, Isolation And Purification, Metabolism, Nuclear Magnetic Resonance, Drug Discovery, Endothelial Cells, Biomolecular, Vascular Endothelial Growth Factor A, Vascular Endothelial Growth Factor Receptor-1,
*** IBB - CNR *** Istituto di Biostrutture e Bioimmagini, CNR, Napoli, Italy.
Vascular endothelial growth factor (VEGF) is a potent angiogenic factor. Its biological activity is mediated by the binding to the extracellular domain of two tyrosine kinase transmembrane receptors: VEGFR1 and VEGFR2. Deletion studies showed that VEGF binding site resides in the first three domains of VEGFR1 and in domains 2 and 3 of VEGFR2. In particular, the second extracellular domain of VEGFR1 (VEGFR1(D2)) contains most of the VEGF binding requirements. Here, we report an efficient expression protocol and the molecular characterization by spectroscopic techniques of VEGFR1(D2). The protein was expressed in E. coli and refolded from inclusion bodies. The recombinant protein assumes the correct fold as assessed by a combination of biochemical and functional assays as well as by NMR characterization. Furthermore, the recombinant VEGFR1(D2) was analyzed by circular dichroism and fluorescence spectroscopy. The protein obtained by this procedure is suitable for the structural characterization of the complexes with receptor binders and to be used in interaction/screening studies. 2010 Wiley Periodicals, Inc.
Aloj L, Aurilio M, Rinaldi V, D'Ambrosio L, Tesauro D, Peitl PK, Maina T, Mansi R, Von Guggenberg E, Joosten L, Sosabowski JK, Breeman WA, De Blois E, Koelewijn S, Melis M, Waser B, Beetschen K, Reubi JC, De Jong M * The EEE project(130 visite) Proc Int Cosm Ray Conf Icrc Universidad Nacional Autonoma De Mexico, 2007; 5(HEPART2): 977-980. Impact Factor:0 DettagliEsporta in BibTeXEsporta in EndNote
351 Records (338 escludendo Abstract e Conferenze). Impact factor totale: 1381.223 (1322.621 escludendo Abstract e Conferenze). Impact factor a 5 anni totale: 1410.156 (1345.443 escludendo Abstract e Conferenze).