Interaction of tetrahydrofolate and other folate derivatives with bacterial sarcosine oxidase.
Sarcosine oxidase from Corynebacterium sp. P-1 binds 2 mol of tetrahydrofolate/mol of enzyme (KD = 8.8 microM). The same stoichiometry is observed with tetrahydropteroyltetraglutamate (KD = 15.4 microM). Binding is also observed with pteroyltetraglutamate and with 5-formyltetrahydrofolate. In the case of the pteroylmonoglutamates, binding appears to be sensitive to changes in the pteridine ring since no binding is observed with 5-methyltetrahydrofolate or with folate. Sarcosine oxidase can be specifically adsorbed onto an affinity matrix prepared by coupling 5-formyltetrahydrofolate to AH-Sepharose. Tetrahydrofolate does not affect the rate of sarcosine oxidation but does block the formation of formaldehyde as a final product. In the presence of tetrahydrofolate, sarcosine oxidation is accompanied by the formation of 5,10-methylenetetrahydrofolate at a rate that exceeds the rate at which formaldehyde (or a precursor) can be released into solution and which is also considerably faster than the nonenzymic reaction of free formaldehyde with tetrahydrofolate. It is suggested that tetrahydrofolate may serve primarily to trap formaldehyde as it is formed at the active site during sarcosine oxidation. The existence of a catalytically significant binding site for tetrahydrofolate appears to be a general property of sarcosine oxidizing enzymes since similar results have previously been obtained with mammalian sarcosine dehydrogenase, an enzyme that is structurally and mechanistically very different from bacterial sarcosine oxidase.
Department of Biological Chemistry, Hahnemann Univeristy School of Medicine, Philadelphia, Pennsylvania 19102.
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D000445:Aldehyde Oxidoreductases / Q000378:metabolism; D003352:Corynebacterium / Q000201:enzymology*; D005492:Folic Acid / Q000378:metabolism; D007700:Kinetics; D010089:Oxidoreductases, N-Demethylating / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D050879:Sarcosine Oxidase; D013329:Structure-Activity Relationship; D013763:Tetrahydrofolates / Q000138:chemical synthesis / Q000378:metabolism*
D013763:Tetrahydrofolates; D005492:Folic Acid; C020145:formaldehyde dehydrogenase (glutathione); D000445:Aldehyde Oxidoreductases; D010089:Oxidoreductases, N-Demethylating; D050879:Sarcosine Oxidase