Genetic Analysis of Glutathione S-transferase A1 Polymorphism in the Chinese Population and the Influence of Genotype on Enzymatic Properties

Abstract

Genetic polymorphisms of human glutathione S-transferases (hGSTs) have important implications for drug efficacy and cancer susceptibility. hGSTA1 is the most abundant subfamily of hGSTs. The aim of this study was to investigate the distribution of hGSTA1 genetic polymorphism in the Chinese population and whether there exists the potential activity alterations caused by this polymorphism. Therefore, genomic DNA was extracted from peripheral blood of 140 healthy Chinese people and 11 normal liver tissues obtained from patients who had undergone liver surgery. Two variants in the promoter region of the hGSTA1 gene were identified by polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP). Activities toward selected substrates of the wild type (hGSTA1*A) and variant (hGSTA1*B) were measured spectrometrically using S-9 fractions from liver samples. Δ⁵-androstene-3,17-dione (AD), cumene hydroperoxide (CuOOH), and 1-chloro-2,4-dinitrobenzene (CDNB) were used as marker substrates toward hGSTA1, hGSTA, and hGST, respectively. The kinetic parameters (K_m, V_max, and V_max/K_m) of hGSTA1 were determined with different concentrations of AD. The results showed that, in the study population, the proportions of hGSTA1*A/*A, hGSTA1*A/*B, and hGSTA1*B/*B genotypes were 75.0, 24.3, and 0.7%, respectively, and the allele frequencies of hGSTA1*A and hGSTA1*B were detected to be 87.1 and 12.9%, respectively. The variant hGSTA1 showed a significantly decreased activity for AD isomerization as compared to the wild type. Kinetic analyses revealed that the V_max value of the variant hGSTA1 was 48% of that of the wild type despite a K_m value of 62% (p < 0.01). This means that the V_max/K_m in the variant hGSTA1 was 76%. These data indicate that the distribution of hGSTA1 gene is polymorphic in Chinese and is different from those in other racial populations. The promoter sequence polymorphism of the hGSTA1 gene is associated with decreased K_m and V_max values of the enzyme in individuals with the variant allele. This variant is also associated with a decrease in hGSTA1 activity toward preferred substrates. This altered activity, however, is accompanied by significant individual variation in the variant population.

glutathione S-transferases, polymorphism, promoter, catalytic activity, enzyme kinetics