We have characterized a second cDNA sequence, pGTH2, for the human liver glutathione S-transferases Ha subunits. It is 95% homologous base-for-base to the Ha subunit 1 cDNA, pGTH1, except for its longer 3' noncoding sequences. Our results indicate that the multiple basic human liver glutathione S-transferases are products of separate genes. The proposal [Kamisaka, K., Habig, W. H., Ketley, J. N., Arias, I. M., and Jakoby, W. B. (1975) Eur. J. Biochem. 60, 153-161] that deamidation may be a physiologically important process for generating glutathione S-transferases isozyme multiplicity can be all but ruled out.