Heterogeneity of glyceraldehyde-3-phosphate dehydrogenase from human brain.
In an attempt to characterize enzymes from human brain capable of dehydrogenating short chain aliphatic aldehydes, four groups of enzymes which catalyze inorganic phosphate-dependent reversible dehydrogenation of glyceraldehyde 3-phosphate as well as short chain aldehydes have been purified and characterized. Three enzyme groups are visualized as multiple bands on isoelectric focusing: E6.6 (pI 6.65, 6.75, 6.85); E6.8 (pI 6.8, 6.9); E8.5 (pI 8.5, 8.6); one enzyme, E9.0, is seen as a single band pI 9.0. The subcellular localization of E6.8, E8.5 and E9.0 appears to be mitochondrial. The mitochondrial enzymes differ slightly in molecular weight: E6.8 is 142,000 with subunits of 36,000 and 38,000; E8.5 is 120,000 with a subunit weight of 29,500; E9.0 is 133,000 with a subunit of 33,000. The E8.5 and E9.0 enzymes also appear to contain Zr as part of their molecular structure. E6.6 (subcellular localization uncertain) is a dimer with a molecular weight of 98,000 and two subunits of 58,000 and 61,000. The specific activity with glyceraldehyde-3-phosphate is: E6.6, 8.6 IU/mg; E6.8, 13 IU/mg; E8.5, 158 IU/mg; E9.0, 620 IU/mg. With glyceraldehyde 3-phosphate and 1,3-diphosphoglyceric acid and Km values of all the enzymes are similar (10-40 microM), except for E6.8 whose Km for glyceraldehyde 3-phosphate is very sensitive to pH and is extremely low at pH 7.0 (2 microM), while being considerably higher than that for the other enzymes at pH 9.0 (170 microM). The molecular properties, Km values as well as high specific activity with glyceraldehyde 3-phosphate identify E6.8, E8.5 and E9.0 as glyceraldehyde-3-phosphate dehydrogenases (EC 1.2.1.12). The catalytic properties of E6.6 are similar to those of E6.8, E8.5 and E9.0, but its molecular properties are different, precluding definite identification.
Ryzlak
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Center of Alcohol Studies, Rutgers University, Piscataway, NJ 08855-0969.
Pietruszko
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D000447:Aldehydes / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002451:Cell Compartmentation; D005986:Glyceraldehyde 3-Phosphate / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007526:Isoelectric Point; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D046911:Macromolecular Substances; D008970:Molecular Weight; D051381:Rats; D013057:Spectrum Analysis; D013379:Substrate Specificity; D015032:Zinc / Q000378:metabolism
D000447:Aldehydes; D007527:Isoenzymes; D046911:Macromolecular Substances; D005986:Glyceraldehyde 3-Phosphate; C005556:propionaldehyde; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D015032:Zinc