Two isoenzymes of glyceraldehyde-3-phosphate dehydrogenase in Caenorhabditis elegans. Isolation, properties, and immunochemical characterization.
Two glyceraldehyde-3-phosphate dehydrogenases have been separated and purified from the nematode Caenorhabditis elegans. As defined by starch gel electrophoresis, the faster-migrating isoenzyme, glyceraldehyde-3-phosphate dehydrogenase-2, increases its activity during postembryonic development. In contrast, the slower-migrating isoenzyme, glyceraldehyde-3-phosphate dehydrogenase-1, is enriched in isolated embryos. Both isoenzymes were initially purified by ammonium sulfate fractionation, gel filtration, and NAD+-agarose affinity chromatography. The separation of both isoenzymes as well as their purification to homogeneity was obtained by preparative chromatofocusing. The subunit molecular weight of each isoenzyme is 38,500 +/- 500. A tetrameric native molecular weight of 157,000 +/- 2000 was determined for glyceraldehyde-3-phosphate dehydrogenase-2. Monospecific rabbit polyclonal antibodies were initially raised against the major isoenzyme and subsequently used to characterize both isoenzymes. Staphylococcus aureas V8 protease digests of each isoenzyme were separated electrophoretically and stained immunochemically, providing evidence that the two isoenzymes differed in their amino acid sequences. Developmental immunocytochemical studies suggest that the embryonic-enriched isoenzyme, glyceraldehyde-3-phosphate dehydrogenase-1, is present in all cells. The second isoenzyme, exhibiting the major activity during postembryonic larval development, may define a body-wall-muscle specific activity which is located within the actin-containing I and A zones of the nematode's sarcomeres.
D000375:Aging; D000818:Animals; D002107:Caenorhabditis / Q000201:enzymology* / Q000254:growth & development; D004625:Embryo, Nonmammalian / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000302:isolation & purification*; D007527:Isoenzymes / Q000302:isolation & purification*; D046911:Macromolecular Substances; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology; D010802:Phylogeny; D013045:Species Specificity
D007527:Isoenzymes; D046911:Macromolecular Substances; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases