Purification and properties of rat lens methionine adenosyltransferase.
Methionine adenosyltransferase (MAT) has been partially purified from rat lenses using a combination of ammonium sulfate fractionation and hydrophobic chromatography on phenyl Sepharose columns. The partially purified enzyme resembles purified Type II MAT from non-hepatic tissues. The Km for methionine is 3.0 microM, and the Km for ATP is 80 microM. The enzyme is activated by potassium ions (25-50 mM), and inhibited by higher concentrations of potassium. A divalent cation (magnesium or manganese) is essential for activity. The Vmax with magnesium is about five times higher than with manganese, but the optimal manganese concentration is around 2.0 mM, compared with 10-20 mM for magnesium. The enzyme is active over a broad pH range, with optimal activity between pH 7.0 and 8.0. The enzyme is inhibited by all three of its products, phosphate, pyrophosphate, and S-adenosylmethionine. Individually phosphate and pyrophosphate are weak inhibitors, but in combination they show a marked synergistic inhibitory effect. Tripolyphosphate is also an effective inhibitor. The inhibition of the enzyme by the cataractogenic agent, dimethylsulfoxide, further confirmed the similarity to Type II MAT.
D000818:Animals; D000906:Antibodies / Q000276:immunology; D004789:Enzyme Activation; D006863:Hydrogen-Ion Concentration; D007908:Lens, Crystalline / Q000201:enzymology*; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008715:Methionine / Q000378:metabolism; D008716:Methionine Adenosyltransferase / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012436:S-Adenosylmethionine / Q000378:metabolism; D014166:Transferases / Q000302:isolation & purification*
D000906:Antibodies; D010710:Phosphates; D008345:Manganese; D012436:S-Adenosylmethionine; D008715:Methionine; D014166:Transferases; D008716:Methionine Adenosyltransferase; D008274:Magnesium; D011188:Potassium