3bto: Difference between revisions

Thiolane 1-oxides are analogs of the carbonyl substrates that bind to the, alcohol dehydrogenase-NADH complex and are potent uncompetitive inhibitors, against alcohol [Chadha, V. K., et al. (1985) J. Med. Chem. 28, 36-40]., The four stereoisomers of 3-butylthiolane 1-oxide (BTO) were separated by, chiral phase chromatography. CD and 1H-NMR spectra identified the, enantiomeric pairs. 1H-NMR chemical shifts were assigned on the basis of, COSY spectra of both diastereoisomers and confirmed by HMQC spectra., Coupling constants were determined through one-dimensional decoupling, experiments. NMR with chiral shift reagents, Eu(hfc)3 [europium tris, [3-[(heptafluoropropyl)hydroxymethylene]-(+)-camphorate]] or, (R)-(-)-N-(3,5-dinitrobenzoyl)-alpha-methylbenzylamine, determined that, the most inhibitory isomer is either 1S,3R or 1R,3S. The chemical shifts, of protons in the thiolane 1-oxide ring were influenced by the whole, structure and were not correlated with the computed Mulliken charges., X-ray crystallography at 2.1 and 1.66 A resolution of the ternary enzyme, complexes with NADH demonstrated that the absolute configuration of the, most inhibitory (Kii = 0.31 microM) stereoisomer is 1S,3R and the next, best inhibitor (Kii = 0.73 microM) is 1S,3S. The thiolane 1-oxide rings, bind in the same position, in the substrate binding site, but the geometry, of the complexes suggests that the sulfoxides are not transition state, analogs. Significantly, the butyl groups of the two isomers are, accommodated differently by flexible amino acid side chains adopting, alternative rotameric conformations.

3BTO is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Equus_caballus Equus caballus] with ZN, NAD and SSB as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Alcohol_dehydrogenase Alcohol dehydrogenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.1 1.1.1.1] Structure known Active Site: NAD. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=3BTO OCA].  

''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov  5 15:27:13 2007''