Serine hydroxymethylase. Specificity of bond cleaveage to form quinonoid intermediates and rate of holoenzyme formation.
L-Serine transhydroxymethylase (5,10-methylenetetrahydrofolate:glycine hydroxymethyltransferase, EC 2.1.2.1) a pyridoxal phosphate-dependent enzyme, has been obtained as a homogeneous preparation with a specific activity of 6.7 mumol benzaldehyde per minute at 30 degrees C at pH 7.5 in N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (Hepes) buffer, with DL-threo-beta-phenylserine as a substrate. This enzyme has been used to study the specificity of bond cleavage in forming quinonoid intermediates from DL and non-asymmetric amino acids. The ability of the generated quinonoids to react with formaldehyde and acetaldehyde has also been studied and evidence obtained for formation of the corresponding beta-hydroxymethyl and beta-hydroxyethyl amino acid derivaties. Apotranshydroxymethylase has been prepared and the rate of holoenzyme formation was found to be 0.52 min-1 by measuring Schiff base formation at 425 nm and 0.66 min-1 as determined from restoration of enzymic activity. A requirement for the presence of mercaptoethanol for complete reactivation was also established by these studies.
D000447:Aldehydes; D000596:Amino Acids; D000818:Animals; D001051:Apoenzymes / Q000378:metabolism; D012696:Glycine Hydroxymethyltransferase / Q000378:metabolism*; D008099:Liver / Q000201:enzymology; D011809:Quinones / Q000378:metabolism*; D012545:Schiff Bases; D012756:Sheep; D014166:Transferases / Q000378:metabolism*
D000447:Aldehydes; D000596:Amino Acids; D001051:Apoenzymes; D011809:Quinones; D012545:Schiff Bases; D014166:Transferases; D012696:Glycine Hydroxymethyltransferase