Mitochondrial GTP-AMP phosphotransferase. 1. Purification and properties.
GTP-AMP phosphotransferase has been purified 116-fold with a yield of 24% from beef heart mitochondria using freeze-thawing, alkali and acid treatment and successive column chromatography on phosphocellulose, Sephadex G-100 and blue-dextran--Sepharose. It has crystallized from poly-(ethylene glycol) and is essential homogeneous by sodium dodecylsulfate electrophoresis and isoelectrofocusing. The specific activity of the crystalline preparation was 290 U/mg. The molecular weight was found to be 26000 and the isoelectric point to be 9.8. Amino acid analysis showed 21 aspartic acid or asparagine, 19 threonine, 12 serine, 26 glutamic acid or glutamine, 15 proline, 16 glycine, 14 alanine, 15 valine, 4 methionine, 12 isoleucine, 28 leucine, 7 tyrosine, 7 phenylalanine, 5 histidine, 14 lysine, 16 arginine, 2 tryptophan, no --SS-- bonds or free --SH. Guanosine(5')pentaphospho(5')adenosine is a very strong inhibitor similar to adenosine(5')pentaphospho(5')adenosine as an inhibitor of cytosolic adenylate kinase.
D000249:Adenosine Monophosphate; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D006160:Guanosine Triphosphate; D008929:Mitochondria, Heart / Q000201:enzymology*; D008970:Molecular Weight; D009703:Nucleoside-Phosphate Kinase; D010770:Phosphotransferases / Q000302:isolation & purification*
D000596:Amino Acids; D000249:Adenosine Monophosphate; D006160:Guanosine Triphosphate; D010770:Phosphotransferases; C021894:nucleoside triphosphate-adenylate kinase; D009703:Nucleoside-Phosphate Kinase