Enzymic properties of different types of human erythrocyte glucose-6-phosphate dehydrogenase, with characterization of two new genetic variants

Abstract

Enzymic properties have been compared in the following five genetic variants of glucose-6-phosphate dehydrogenase from human erythrocytes: the two common variants with normal activity, A and B; the common variant associated with enzyme deficiency, A-; and two new rare variants, “Ijebu-Ode” and “Ita-Bale.” The maximal velocity of the enzyme reaction (V_max) increases steadily with pH over the entire range explored (from pH 5.5 to 9.5) for all enzyme variants when buffers are used that show no specific ion effects on enzyme activity. Small differences are found among the variants in the pH range 7.5-8.2, where A and B show a “peak and trough,” while A-, “Ijebu-Ode,” and “Ita-Bale” exhibit a plateau. When the effects of reagents that bind to sulphydryl groups are compared, iodoacetate, bromoacetate, and iodoacetamide are weak inhibitors, while N-ethylmaleimide (NEM) and hydroxymercuribenzoate (HMB) are potent inhibitors. The last two reagents have differential inhibitory action on different variants; one of these, “Ijebu-Ode,” is strikingly resistant to HMB and totally resistant to NEM (up to 3 mmoles/liter). The enzyme inactivation as a function of temperature exhibits distinctive profiles for all variants examined. Both of the new variants described differ significantly from the normal B type in several respects: “Ijebu-Ode” in electrophoretic mobility, thermostability, dependence of V_max on pH, and resistance to sulphydryl group reagents; “Ita-Bale” in electrophoretic mobility, Michaelis constant (K_m) for glucose-6-phosphate, and dependence of V_max on pH. When these data are compared with those available in the literature, both variants are different from all those previously described. The estimated frequencies of the corresponding genes in western Nigeria are between 0.0005 and 0.0025 for “Ijebu-Ode” and less than 0.0005 for “Ita-Bale”. The A- variant, compared to A, has a distinctly higher K_m for 2-deoxyglucose-6-phosphate and is more inhibited by very low concentrations of HMB. These are the first observed differences in kinetic properties between A and A-.