Random mutagenesis of human serine racemase reveals residues important for the enzymatic activity

0343324 - ÚOCHB 2011 RIV CZ eng J - Článek v odborném periodiku
Hoffman, Hillary Elizabeth - Jirásková, Jana - Zvelebil, M. - Konvalinka, Jan
Random mutagenesis of human serine racemase reveals residues important for the enzymatic activity.
Collection of Czechoslovak Chemical Communications. Roč. 75, č. 1 (2010), s. 59-79. ISSN 0010-0765
Grant CEP: GA MŠMT 1M0508
Výzkumný záměr: CEZ:AV0Z40550506
Klíčová slova: D-serine * serine racemase * random mutagenesis
Kód oboru RIV: CE - Biochemie
Impakt faktor: 0.853, rok: 2010

Human serine racemase (hSR) is a cytosolic pyridoxal-5′-phosphate dependent enzyme responsible for production of D-serine in the central nervous system. D-Serine acts as an endogenous coagonist of N-methyl-D-aspartate receptor ion channels. Increased levels of D-serine have been linked to amyotrophic lateral sclerosis and Alzheimer’s disease.Here, we present a strategy for the generation and screening of random hSR mutants. From a library of randomly mutated hSR variants, twenty-seven soluble mutants were selected, expressed, and evaluated for enzymatic activity. Taking three carefully characterized mutants as an example, we show how this strategy can be used to pinpoint structurally and functionally important residues. In particular, we identify S84 and P111 as residues crucial for hSR activity and C217 and K221 as residues important for binding of the Mg2+ cofactor.
Trvalý link: http://hdl.handle.net/11104/0185830