Identification of inhibitors of the E. coli cyclopropane fatty acid synthase from the screening of a chemical library: In vitro and in vivo studies. - Université Clermont Auvergne Accéder directement au contenu
Article Dans Une Revue Biochimica et Biophysica Acta - Molecular Cell Research Année : 2008

Identification of inhibitors of the E. coli cyclopropane fatty acid synthase from the screening of a chemical library: In vitro and in vivo studies.

Dominique Guianvarc'H (1) , Guangqi E (2) , Thierry Drujon (1) , Camille Rey (3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14) , Qian Wang (15, 16, 17, 18, 19, 20, 21, 22, 23) , Olivier Ploux (1)

Résumé

Using an automated coupled colorimetric assay for the Escherichia coli cyclopropane fatty acid synthase (CFAS), we have screened an academic chemical library of 3040 compounds, to identify new inhibitors of this enzyme. We identified 8 compounds as potent inhibitors of this enzyme, with IC(50) ranging from 1 to 10 microM, in the presence of 750 microM S-adenosyl-l-methionine and 1 mg/mL phospholipids. We conducted kinetic analyses of the inhibition of the CFAS using dioctylamine and three inhibitors identified in this report: sinefungin, 1, a synthetic S-adenosyl-l-homocysteine analog, 2, and an indoloquinolizine derivative, 3. The inhibition patterns observed were interpreted assuming that the E. coli CFAS operated via an ordered Bi Bi mechanism with binding of S-adenosyl-l-methionine first. Dioctylamine was the most potent inhibitor with a competitive inhibition constant of 130 nM with respect to the phospholipids. Compound 2 bound to the two substrate-binding sites of the enzyme suggesting that it acted as a bisubstrate analog (apparent inhibition constant, K(I)=6 microM). Compound 2 was also found to completely inhibit cyclopropanation of the phospholipids in growing E. coli cells, at 150 microM. This molecule is thus the first inhibitor of a cyclopropane synthase that is active in vivo, contrary to sinefungin and other analogs that are only active on the isolated enzyme.

Dates et versions

hal-00294830 , version 1 (10-07-2008)

Identifiants

Citer

Dominique Guianvarc'H, Guangqi E, Thierry Drujon, Camille Rey, Qian Wang, et al.. Identification of inhibitors of the E. coli cyclopropane fatty acid synthase from the screening of a chemical library: In vitro and in vivo studies.. Biochimica et Biophysica Acta - Molecular Cell Research, 2008, 1764 (11), pp.1381-1388. ⟨10.1016/j.bbapap.2008.04.019⟩. ⟨hal-00294830⟩
464 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More