Purification and properties of streptococcal nicotinamide adenine dinucleotide glycohydrolase.
Highly purified streptococcal nicotinamide adenine dinucleotide glycohydrolase (NADase) was obtained by utilizing disodium tetrathionate to protect the enzyme by blocking the sulfhydryl groups of streptococcal proteinase. This was followed by two-step ion-exchange chromatography. The pure enzyme, demonstrated as a single band on sodium dodecyl sulfate/polyacrylamide gel electrophoresis, had a specific activity of 11,200 NADase units per mg of protein and was devoid of hemolytic activity. NADase had a molecular weight of about 55,000 as determined by gel filtration, by summation of amino acid residues, and by sodium dodecyl sulfate/gel electrophoresis. The purified enzyme had optimal activity at pH 7.3 and at 40 C and a calculated Km of 5.1 times 10- minus 4 mM. It was inhibited by alpha-iodoacetamide.
D000596:Amino Acids / Q000032:analysis; D000645:Ammonium Sulfate; D011232:Chemical Precipitation; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors; D006863:Hydrogen-Ion Concentration; D007460:Iodoacetamide / Q000494:pharmacology; D008970:Molecular Weight; D009699:N-Glycosyl Hydrolases; D009244:NAD+ Nucleosidase / Q000302:isolation & purification / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D013297:Streptococcus pyogenes / Q000201:enzymology*; D013301:Streptolysins / Q000378:metabolism; D013696:Temperature; D013776:Tetrathionic Acid
D000596:Amino Acids; D004791:Enzyme Inhibitors; D013301:Streptolysins; D013776:Tetrathionic Acid; D009699:N-Glycosyl Hydrolases; D009244:NAD+ Nucleosidase; D010447:Peptide Hydrolases; D000645:Ammonium Sulfate; D007460:Iodoacetamide