Lj. Dickmann et al., Identification and functional characterization of a new CYP2C9 variant (CYP2C9*5) expressed among African Americans, MOLEC PHARM, 60(2), 2001, pp. 382-387
CYP2C9 is a polymorphic gene for which there are four known allelic variant
s; CYP2C9*1, CYP2C9*2, CYP2C9*3, and CYP2C9*4. In the present study, DNA fr
om 140 European Americans and 120 African Americans was examined by single-
strand conformational polymorphism and restriction fragment length polymorp
hism analyses, resulting in the identification of a new CYP2C9 variant, CYP
2C9*5. This variant is derived from a C1080G transversion in exon 7 of CYP2
C9 that leads to an Asp360Glu substitution in the encoded protein. The CYP2
C9*5 variant was found to be expressed only in African Americans, such that
approximately 3% of this population carries the CYP2C9*5 allele. The varia
nt expressed in, and purified from, insect cells infected with a recombinan
t baculovirus. Comparative kinetic studies using the purified wild-type pro
tein CYP2C9*1; the IIe359Leu variant, CYP2C9*3; and the Asp 360Glu variant
CYP2C9*5 were carried out using (S)-warfarin, diclofenac, and lauric acid a
s substrates. The major effect of the Asp360Glu mutation was to increase th
e K-m value relative to that of CYP2C9*1 for all three substrates: 12-fold
higher for (S)-warfarin 7-hydroxlation, 5-fold higher for the 4'-hydroxylat
ion of ldiclofnac and 3-fold higher for the omega -1 hydroxlation of lauric
acid, V-max values differed less that K-m values between the CYp2C9*1 and
CYP2C9*5 proteins. In vitro intrinsic clearances for CYP2C9*5 calculated as
the ratio of V-max/K-m, ranged fro 8 to 18% of CYP2C9*1 values. The corres
ponding ratio for CYP2C9*3 was 4 to 13%. Accordingly, the in vitro data sug
gest that carriers of the CYP2C9*5 allele would eliminate CYP2C9 substrates
at slower rates relative to person expressing the wild-type protein.