An improved rapid assay for S-adenosyl-L-homocysteine hydrolase.
A coupled enzyme system was devised to assay S-adenosyl-L-homocysteine hydrolase activity spectrophotometrically and to stain the hydrolase selectively in polyacrylamide gels. The assay procedure monitors the formation of uric acid derived from the catabolism of the adenosine moiety of S-adenosylhomocysteine. The staining procedure allows the determination of the molecular weight of the hydrolase when extracts are electrophoresed on polyacrylamide gradient gels and compared to standard of known molecular weight. The specificity of the enzyme for the homocysteine moiety was also investigated by testing modified compounds as substrates. All the analogs tested were inactive as substrates for hydrolysis, indicating a strict specificity.
D043383:Adenosylhomocysteinase; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005260:Female; D006867:Hydrolases / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D051381:Rats; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013379:Substrate Specificity
D006867:Hydrolases; D043383:Adenosylhomocysteinase