The sodium pump glutaconyl-CoA decarboxylase from Acidaminococcus fermentans. Specific cleavage by n-alkanols.
Glutaconyl-CoA decarboxylase from Acidaminococcus fermentans was inactivated by incubation with n-alkanols at 37 degrees C. The concentration of the alcohol required for complete inactivation decreased with increasing chain length; e.g. 2 M ethanol was as potent as 2 mM hexanol or 0.5 mM decanol. The data indicate a binding of the alcohol to the enzyme with an energy of about 4 kJ/methylene group. Sodium ions prevented the inactivation (50% at 30 mM NaCl). K+, NH4+, Cs+ and Mg2+ had no influence, whereas Li+ was ten times less effective than Na+. The enzyme was cleaved during the inactivation into a soluble part, consisting of the alpha (Mr 120,000) and beta polypeptide chains (60,000), whereas the hydrophobic gamma chain (30,000) precipitated. The soluble part catalysed the sodium-ion-independent but avidin-sensitive glutaconyl-CoA/crotonyl-CoA exchange as measured with the substrates [3-3H]crotonyl-CoA and unlabelled glutaconate and with glutaconate CoA-transferase as auxiliary enzyme. In the presence of free biotin or its methyl ester the soluble part catalysed the formation of crotonyl-CoA from glutaconyl-CoA (apparent Km for biotin 40 mM, Vmax 1% of the native decarboxylation reaction). This apparent reactivation was most likely caused by the carboxylation of free biotin. Based on these and other observations the following functions may be assigned to the different polypeptide chains of glutaconyl-CoA decarboxylase: biotin carrier (alpha), carboxytransferase (beta) and carboxylase, the actual sodium pump (gamma).
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D000438:Alcohols / Q000494:pharmacology*; D000596:Amino Acids / Q000032:analysis; D001710:Biotin / Q000494:pharmacology; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004793:Enzyme Reactivators; D007473:Ion Channels / Q000378:metabolism*; D007700:Kinetics; D012964:Sodium / Q000378:metabolism* / Q000494:pharmacology; D045854:Veillonellaceae / Q000201:enzymology*
D000438:Alcohols; D000596:Amino Acids; D004793:Enzyme Reactivators; D007473:Ion Channels; D001710:Biotin; D012964:Sodium; D002262:Carboxy-Lyases; C023060:glutaconyl CoA decarboxylase