title: Guanidinoacetate methyltransferase activity in tissues and cultured cells.
abstract: Guanidinoacetate methyltransferase, the enzyme catalyzing the last step in creatine biosynthesis, has previously been considered to be restricted to a few tissues, but it has been found to occur in the cultured cells H4Az C2 rat hepatoma, N4TG1 mouse neuroblastoma, and IMR-90 human fetal lung fibroblast, as well as in skeletal and cardiac muscle of the rat. Activity was highest in the hepatoma, but tissues and cultured cells of nonhepatic origin had 5-20% of the activity of rat liver. Dialyzed 100,000g supernatants prepared from cultured cells or skeletal muscle tissue yielded values for apparent Km in the range of 1.2-3.4 microM for S-adenosylmethionine and 0.050-0.096 mM for guanidinoacetate. Intact monolayers of the three types of cultured cells converted labeled guanidinoacetate in the culture medium to creatine, which was identified by chromatographic behavior and by reaction with creatine kinase. The amounts of guanidinoacetate converted to creatine by fibroblasts and neuroblastoma cells during an 18-h period of incubation suggested that synthesis was proceeding at rates approaching Vmax, even in medium containing the relatively low physiological concentrations of guanidinoacetate. Fibroblast and neuroblastoma cell monolayers also have the capacity to take up creatine provided in the culture medium. The amounts of creatine taken up by monolayers of those cells were measured under the same conditions that were used for measurement of creatine synthesis. Comparison of the amounts of creatine synthesized with the amounts taken up showed that synthesis can make a significant contribution to intracellular pools of creatine plus phosphocreatine in fibroblasts and neuroblastoma cells.
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003401:Creatine / Q000096:biosynthesis / Q000378:metabolism; D003402:Creatine Kinase / Q000378:metabolism; D005333:Fetus; D005347:Fibroblasts / Q000201:enzymology; D050937:Guanidinoacetate N-Methyltransferase; D006801:Humans; D008114:Liver Neoplasms, Experimental / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008297:Male; D008780:Methyltransferases / Q000302:isolation & purification*; D009132:Muscles / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology; D009447:Neuroblastoma / Q000201:enzymology; D051381:Rats
chemical_list: D008780:Methyltransferases; C494508:Gamt protein, rat; D050937:Guanidinoacetate N-Methyltransferase; D003402:Creatine Kinase; D003401:Creatine