title: Allosteric regulation of mouse brain serine racemase.
abstract: Serine racemase, purified from mouse brain, consisted of two isoforms. They had similar enzymatic properties and had molecular weights of about 55 kDa based on size exclusion chromatography. This is about twice that reported from its electrophoretic mobility on SDS gels or from the amino acid sequence of the recombinant enzyme. In addition to the previously reported requirements for pyridoxal phosphate and reducing agents, we found that both forms of the enzyme required Mg2+ and were strongly stimulated by yeast extract. The yeast extract could be replaced by ATP, GTP, or ADP and, to a lesser extent, by other nucleotides. In the presence of 1 mM ATP, the Km for L-serine decreased from 13 mM to 1.8 mM with little change in Vmax, indicating an allosteric mechanism for nucleotide activation. In addition to acting as a serine racemase, the enzyme has been reported to act on L-serine O-sulfate as a dehydratase. When measured by HPLC, after derivatization with 2,4 dinitrophenylhydrazine, we found, as expected, a very rapid formation of pyruvate from this substrate. L-serine was also converted to pyruvate at about twice the racemization rate. L-serine O-sulfate dehydration was inhibited by ATP, while L-serine dehydration, like racemization, was activated by nucleotides, indicating that, for L-serine, dehydration and racemization take place at the same site.
authors.affiliation: Division of Neurochemistry, Nathan Kline Institute for Psychiatric Research, Orangeburg, New York 10962, USA.
authors.forename: Amos
authors.initials: A
authors.lastname: Neidle
authors.forename: David S
authors.initials: DS
authors.lastname: Dunlop
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D051379:Mice; D019998:Racemases and Epimerases / Q000302:isolation & purification / Q000378:metabolism*; D000099300:Serine Racemase
chemical_list: D019998:Racemases and Epimerases; D000099300:Serine Racemase