In searching the expressed sequence tag (EST) data-base of GenBank with coding sequences of 11 known human glutathione S-transferases in conjunction with bioinformatic analysis, we have identified five ESTs that encode a new human glutathione S-transferase (GST) designated GST A4. The cDNA clone (I.M.A.G.E. Consortium cDNA Clone ID 515157) had an insert length of 1279 bp and contains an open reading frame of 666 bp, which encodes a protein of 222 amino acid residues. The GST A4 protein is identical in length to human GST A1 and A2 and is 54% identical to human GST A1 and A2. Sequence comparison with other human GSTs suggests that it is a new GST belonging to the alpha class GSTs. Northern blot analysis and EST database searches have demonstrated that the GST A4 mRNA is expressed at a high level in brain, placenta, and skeletal muscle and much lower in lung and liver. Analysis of the sequence tagged site (STS) database indicated that the GST A4 gene is located on chromosome 6. This STS represents a previously unidentified transcript further confirming the novelty of the new sequence.