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Random mutagenesis of human serine racemase reveals residues important for the enzymatic activity
- 1.0343324 - ÚOCHB 2011 RIV CZ eng J - Journal Article
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
R&D Projects: GA MŠMT 1M0508
Institutional research plan: CEZ:AV0Z40550506
Keywords : D-serine * serine racemase * random mutagenesis
Subject RIV: CE - Biochemistry
Impact factor: 0.853, year: 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.
Permanent Link: http://hdl.handle.net/11104/0185830
Number of the records: 1