caa a22 4500 475744497 CHVBK 20180406123509.0 cr unu---uuuuu 170329e20000701xx s 000 0 eng 10.1023/A:1026439531005 doi (NATIONALLICENCE)springer-10.1023/A:1026439531005 Characterization of Glyoxalase I (E. coli)—Inhibitor Interactions by Electrospray Time-of-Flight Mass Spectrometry and Enzyme Kinetic Analysis [Elektronische Daten] [Ellen Stokvis, Susan Clugston, John Honek, Albert Heck] Potential inhibitors of the enzyme glyoxalase I from Escherichia coli have been evaluated using a combination of electrospray mass spectrometry and conventional kinetic analysis. An 11-membered library of potential inhibitors included a glutathione analogue resembling the transition-state intermediate in the glyoxalase I catalysis, several alkyl-glutathione, and one flavonoid. The E. coli glyoxalase I quaternary structure was found to be predominantly dimeric, as is the homologous human glyoxalase I. Binding studies by electrospray revealed that inhibitors bind exclusively to the dimeric form of glyoxalase I. Two specific binding sites were observed per dimer. The transition-state analogue was found to have the highest binding affinity, followed by a newly identified inhibitor; S-{2-[3-hexyloxybenzoyl]-vinyl}glutathione. Kinetic analysis confirmed that the order of affinity established by mass spectrometry could be correlated to inhibitory effects on the enzymatic reaction. This study shows that selective inhibitors may exist for the E. coli homologue of the glyoxalase I enzyme. Plenum Publishing Corporation, 2000 Glyoxalase I nationallicence bioaffinity mass spectrometry nationallicence noncovalent interactions nationallicence inhibitor screening nationallicence Stokvis Ellen Department of Biomolecular Mass Spectrometry, Bijvoet Center for Biomolecular Research, and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands aut Clugston Susan Department of Chemistry, University of Waterloo, N2L 3G1, Waterloo, Ontario, Canada aut Honek John Department of Chemistry, University of Waterloo, N2L 3G1, Waterloo, Ontario, Canada aut Heck Albert Department of Biomolecular Mass Spectrometry, Bijvoet Center for Biomolecular Research, and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/5(2000-07-01), 389-397 0277-8033 19:5<389 2000 19 10930 https://doi.org/10.1023/A:1026439531005 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1026439531005 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Stokvis Ellen Department of Biomolecular Mass Spectrometry, Bijvoet Center for Biomolecular Research, and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands aut NATIONALLICENCE 700 1- Clugston Susan Department of Chemistry, University of Waterloo, N2L 3G1, Waterloo, Ontario, Canada aut NATIONALLICENCE 700 1- Honek John Department of Chemistry, University of Waterloo, N2L 3G1, Waterloo, Ontario, Canada aut NATIONALLICENCE 700 1- Heck Albert Department of Biomolecular Mass Spectrometry, Bijvoet Center for Biomolecular Research, and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/5(2000-07-01), 389-397 0277-8033 19:5<389 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer