RT Journal Article
SR Electronic
T1 FUNCTIONAL SIGNIFICANCE OF A NATURAL ALLELIC VARIANT OF HUMAN CARBONYL REDUCTASE 3 (<em>CBR3</em>)
JF Drug Metabolism and Disposition
JO Drug Metab Dispos
FD American Society for Pharmacology and Experimental Therapeutics
SP 254
OP 257
DO 10.1124/dmd.104.002006
VO 33
IS 2
A1 Lakhman, Sukhwinder S.
A1 Ghosh, Debashis
A1 Blanco, Javier G.
YR 2005
UL http://dmd.aspetjournals.org/content/33/2/254.abstract
AB Human carbonyl reductase (CBR) activity accounts for a significant fraction of the metabolism of endogenous and xenobiotic carbonyl compounds. It is possible that genetic polymorphisms in CBR1 and CBR3 are key for the wide interindividual variability in the disposition of CBR drug substrates. We pinpointed a single nucleotide polymorphism in CBR3 (CBR3 V244M) that encodes for a V244 to M244 change. Blacks showed a higher frequency of the M244 allele (q = 0.51, n = 49) than did whites (q = 0.31, n = 70; p = 0.003). In addition, DNA variation panels from 10 ethnic groups presented a wide range of CBR3 V244M genotype distributions. Kinetic experiments with the recombinant CBR3 protein variants and menadione revealed that CBR3 M244 has significantly higher Vmax than does CBR3 V244 (Vmax CBR3 M244 = 40.6 ± 1.3 μmol/min · mg versus Vmax CBR3 V244 = 19.6 ± 2.0 μmol/min · mg, p = 0.002). In contrast, both isoforms presented similar Km values (Km CBR3 M244 = 22.9 ± 2.9 μM versus Km CBR3 V244 = 24.6 ± 3.2 μM, p = 0.43). Assays with NADP(H) demonstrated a higher VmaxNADP(H) (1.6-fold) and increased catalytic efficiency (VmaxNADP(H)/KmNADP(H)) for CBR3 M244 compared with CBR3 V244 (p = 0.013). Comparative three-dimensional analyses based on the structure of the homologous porcine carbonyl reductase suggested that the V244M substitution is positioned in a region critical for interactions with the NADP(H) cofactor. These studies demonstrate that the common CBR3 V244M polymorphism encodes for CBR3 isoforms with distinctive enzymatic properties. The American Society for Pharmacology and Experimental Therapeutics