Worm Breeder's Gazette 7(2): 29b
These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.
We are studying the developmental regulation of two isoenzymes of glyceraldehyde-3-phosphate dehydrogenase (G3PDH) in the small nematode, Caenorhabditis ned by starch gel electrophoresis, the embryonic specific isoenzyme, G3PDH-1, disappears during post-embryonic development. In contrast, the faster migrating isoenzyme, G3PDH-2, increases its activity during post-embryogenesis. In order to elucidate the molecular mechanism(s) underlying the temporal and/or spatial differential expression of G3PDH-1 and G3PDH-2 we have optimized the following G3PDH purification scheme. Both enzymes were purified to near homogeneity by ammonium sulfate fractionation, gel filtration and NAD-agarose affinity chromatography. Complete homogeneity as well as the separation of both isoenzymes was achieved by preparative 'chromatofocusing.' Both isoenzymes exhibited identical native and subunit molecular weights of 131,000 and 38,500, respectively. We believe that these two isoenzymes are encoded by separate genes because opposite mobilities were observed for these two isoenzymes in the related nematode, Caenorhabditis briggsae. To confirm this hypothesis, partial peptide analysis of each purified isoenzyme in Caenorhabditis conducted.