Complex, unusual conformational changes in kidney betaine aldehyde dehydrogenase suggested by chemical modification with disulfiram.
The NAD+-dependent animal betaine aldehyde dehydrogenases participate in the biosynthesis of glycine betaine and carnitine, as well as in polyamines catabolism. We studied the kinetics of inactivation of the porcine kidney enzyme (pkBADH) by the drug disulfiram, a thiol-reagent, with the double aim of exploring the enzyme dynamics and investigating whether it could be an in vivo target of disulfiram. Both inactivation by disulfiram and reactivation by reductants were biphasic processes with equal limiting amplitudes. Under certain conditions half of the enzyme activity became resistant to disulfiram inactivation. NAD+ protected almost 100% at 10 microM but only 50% at 5mM, and vice versa if the enzyme was pre-incubated with NAD+ before the chemical modification. NADH, betaine aldehyde, and glycine betaine also afforded greater protection after pre-incubation with the enzyme than without pre-incubation. Together, these findings suggest two kinds of active sites in this seemingly homotetrameric enzyme, and complex, unusual ligand-induced conformational changes. In addition, they indicate that, in vivo, pkBADH is most likely protected against disulfiram inactivation.
Ayala-Castro
Hector G
HG
Coordinación de Ciencia de los Alimentos, Centro de Investigación en Alimentación y Desarrollo A.C., Hermosillo, Sonora 83100, Mexico.
Valenzuela-Soto
Elisa M
EM
Figueroa-Soto
Ciria G
CG
Muñoz-Clares
Rosario A
RA
D000444:Aldehyde Dehydrogenase / Q000737:chemistry* / Q000648:ultrastructure*; D000818:Animals; D050728:Betaine-Aldehyde Dehydrogenase / Q000737:chemistry* / Q000648:ultrastructure*; D003198:Computer Simulation; D004221:Disulfiram / Q000737:chemistry*; D004795:Enzyme Stability; D007668:Kidney / Q000201:enzymology*; D008956:Models, Chemical; D008958:Models, Molecular; D011487:Protein Conformation; D013552:Swine
D000444:Aldehyde Dehydrogenase; D050728:Betaine-Aldehyde Dehydrogenase; D004221:Disulfiram