SEPARATION BY FPLC CHROMATOFOCUSING OF UDP-GLUCOSYLTRANSFERASES FROM 3 DEVELOPMENTAL STAGES OF DROSOPHILA-MELANOGASTER

Variation of UDP-glucosyltransferase activity, during Drosophila melan ogaster development, was analyzed. The endogenous metabolite xanthuren ic acid and the xenobiotic compounds 1-naphthol and 2-naphthol were us ed as substrates. Developmentally regulated differences were observed for the three substrates, suggesting the presence of UDP-glucosyltrans ferase isoenzymes. This was further confirmed by FPLC chromatofocusing on a Mono P column: seven peaks of UDP-glucosyltransferase activity ( pHs: greater than or equal to 6.3, 5.8, 5.5, 4.9, 4.5, 4.2, less than or equal to 4.0) with either single or overlapping substrate specifici ty were detected. A single xanthurenic acid:UDP-glucosyltransferase ac tivity (pl 5.8) was found throughout development. In contrast, a gradu al increase in the number of 2-napthol:UDP-glucosyltransferase isoenzy mes (pl from 6.3 to 4.0) was observed during development, whereas no i soenzymes specific for 1-naphthol were resolved. Based on the distribu tion and substrate specificity of the eluted peaks in the three develo pmental stages analyzed, the presence of seven or possibly eight UDP-g lucosyltransferase isoenzymes is proposed. (C) 1997 Wiley-Liss, Inc.