Study of non-covalent enzyme-inhibitor complexes of aldose reductase by electrospray mass spectrometry.
Specific non-covalent interactions between aldose reductase (AR), its NADP+ cofactor and five inhibitors have been characterized by electrospray mass spectrometry (ES-MS). These results indicated that the protein could be desorbed and maintained in the gas phase in a form very close to its native conformation. Collisionally induced dissociation (CID)-MS and CID-MS-MS showed that the adenosine diphosphate part of the cofactor interacts strongly with AR. The relative stability of the ternary AR x NADP+ x inhibitor complexes was established and successfully correlated with the IC50 values. All inhibitors were shown to only bind to AR holoenzyme. These results are important for the field of drug development insofar as ES-MS might provide a rapid and very sensitive method for the screening of potential drugs or for the identification of compounds displaying high binding affinity to a target biomolecule.
Potier
N
N
Laboratoire de Spectrométrie de Masse Bio-Organique, URA 31, CNRS-Université Louis Pasteur, Faculté de chimie, Strasbourg, France.
Barth
P
P
Tritsch
D
D
Biellmann
J F
JF
Van Dorsselaer
A
A
D000085:Acetates / Q000494:pharmacology; D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors*; D000818:Animals; D001051:Apoenzymes / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000737:chemistry*; D005663:Furans / Q000494:pharmacology; D007093:Imidazoles / Q000737:chemistry; D048289:Imidazolidines; D007908:Lens, Crystalline / Q000201:enzymology; D013058:Mass Spectrometry / Q000379:methods*; D009249:NADP / Q000737:chemistry; D009281:Naphthalenes / Q000737:chemistry; D012236:Rhodanine / Q000031:analogs & derivatives / Q000494:pharmacology; D013552:Swine; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines
D000085:Acetates; D001051:Apoenzymes; D004791:Enzyme Inhibitors; D005663:Furans; D007093:Imidazoles; D048289:Imidazolidines; D009281:Naphthalenes; C104583:ONO 1; C104584:ONO 3; D013844:Thiazoles; D053778:Thiazolidines; C040550:tolrestat; C038131:epalrestat; D009249:NADP; D012236:Rhodanine; D000449:Aldehyde Reductase; C026411:sorbinil