ABSTRACT
1. The amino acid sequence of D-glyceraldehyde-3-phosphate dehydrogenase from the extreme thermophile Thermus aquaticus has been elucidated. 2. The polypeptide contains 332 amino acids and its sequence is 70% identical with that of the enzyme from the moderate thermophile Bacillus stearothermophilus. 3. In contrast to less thermostable forms of the enzymes from B. stearothermophilus, pig, lobster and yeast, the T. aquaticus enzyme has only one cysteine residue, namely cysteine-149 which is required for catalysis.
Subject(s)
Glyceraldehyde-3-Phosphate Dehydrogenases/analysis , Thermus/enzymology , Amino Acid Sequence , Amino Acids/analysis , Animals , Cyanogen Bromide , Cysteine/analysis , Geobacillus stearothermophilus/enzymology , Nephropidae/enzymology , Pepsin A , Peptide Fragments/analysis , Swine/metabolism , Trypsin , Yeasts/enzymologyABSTRACT
1. D-Glyceraldehyde-3-phosphate dehydrogenase from two thermophilic bacteria has been purified by procedures including affinity chromatography on NAD+-Sepharose. 2. Methods for making NAD+-free enzyme are also described. 3. Both the holo and apo forms of the enzyme from Bacillus stearothermophilus have been crystallised. 4. The enzymes are tetrameric and composed of four chemically identical polypeptide chains of molecular weight 36,000. 5. The enzymes are much more stable to heat than their counterparts from mesophiles.
Subject(s)
Geobacillus stearothermophilus/enzymology , Glyceraldehyde-3-Phosphate Dehydrogenases/isolation & purification , Thermus/enzymology , Chromatography, Affinity , Crystallization , Glyceraldehyde-3-Phosphate Dehydrogenases/metabolism , Kinetics , Molecular Weight , NAD/metabolism , TemperatureSubject(s)
Geobacillus stearothermophilus/enzymology , Glyceraldehyde-3-Phosphate Dehydrogenases , Gram-Negative Aerobic Bacteria/enzymology , Amino Acid Sequence , Glyceraldehyde-3-Phosphate Dehydrogenases/analysis , Glyceraldehyde-3-Phosphate Dehydrogenases/isolation & purification , Glyceraldehyde-3-Phosphate Dehydrogenases/metabolism , Guanidines/pharmacology , Hot Temperature , Molecular Weight , Muscles/enzymology , NAD/analysis , Protein Denaturation/drug effectsSubject(s)
Bacteria/enzymology , Glyceraldehyde-3-Phosphate Dehydrogenases/isolation & purification , Chromatography , Chromatography, Affinity , Chromatography, DEAE-Cellulose , Deoxyribonucleases , Drug Stability , Electrophoresis, Polyacrylamide Gel , Glyceraldehyde-3-Phosphate Dehydrogenases/metabolism , Hot Temperature , Hydroxyapatites , Molecular Weight , Muscles/enzymology , NAD , Polysaccharides , Sodium Dodecyl Sulfate , Species Specificity , ThermodynamicsABSTRACT
A disease of Angus cattle previously known as pseudolipidosis has been shown to be an inherited lysosomal storage disease, in which an oligosaccharide containing mannose and glucosamine is the storage substance. Diseased animals have a near-absolute deficiency of the lysosomal enzyme, alpha-mannosidase, whereas heterozygotes have a partial deficiency of this enzyme. The condition is analogous to the human disease known as mannosidosis.
