Abstract

Specific cDNA probes and antisera were employed to interpret genetic polymorphisms of human Mu-class glutathione S-transferases and to provide a basis for identifying individual forms in human tissues. A cDNA probe that cross-hybridized with various human and rodent Mu-glutathione S-transferase transcripts, hybridized with at least three discrete components by Northern analysis of RNA from human tissue. The smallest (1.3 kb) transcript was identified as the one that encodes GSTM3-3 subunits. A form designated GSTM5, was cloned from a human brain cDNA library and its sequence determined. The open reading frame of GSTM5 shared a high degree of homology with the sequences of other Mu-class glutathione S-transferases, but its 846-nucleotide 3'-noncoding region was unique and considerably larger than that of any of the other Mu forms. Specific synthetic peptide antigens were utilized to distinguish among Mu-class glutathione S-transferases in different tissues of representative individuals. The primary hepatic transcript was that encoding GSTM1-1 with much lesser amounts of GSTM3-3, but livers were devoid of GSTM2-2, and GSTM5-5. Immunoblots confirmed that null-phenotype individuals lacked the GSTM1 gene rather than its GSTM2 homologue that is nearly identical in its exon sequences. The null phenotype therefore was conspicuous in liver, where GSTM1-1 ordinarily was the predominant Mu transcript, but brain and testis contained all four forms. A general strategy was devised to distinguish among and assign primary structures to individual glutathione S-transferases from human tissue.