Mechanism of eukaryotic serine racemase-catalyzed serine dehydration

Ito, Tomokazu; Matsuoka, Mai; Goto, Masaru; Watanabe, Soichiro; Mizobuchi, Taichi; Matsushita, Kazuma; Nasu, Ryoma; Hemmi, Hisashi; Yoshimura, Teizo

doi:10.1016/j.bbapap.2020.140460

Cited by 3 publications

(2 citation statements)

References 25 publications

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“…D-serine dehydratase from S. cerevisiae was overexpressed as an N-terminal His-tagged protein in E. coli KRX strain (Promega) harboring pET-15b-Dsd1p and purified homogeneity as described previously (Ito et al 2008). SR from mouse (N-terminal His-tag) was overexpressed in E. coli Rosetta2 (DE3) harboring pET-28a-mSR and purified as previously described [25]. E. coli asparaginase (AnsA) or asparagine synthase (AsnA) was overexpressed as an N-terminal His-tagged protein with strain from the ASKA collection (National BioResource Project (NIG,Japan): E.coli) [26].…”

Section: Enzyme Purificationmentioning

confidence: 99%

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Urinary l-erythro-β-hydroxyasparagine—a novel serine racemase inhibitor and substrate of the Zn2+-dependent d-serine dehydratase

et al. 2021

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In this study, we identified L-erythro-β-hydroxyasparagine (L-β-EHAsn) found abundantly in human urine, as a novel substrate of Zn2+-dependent D-serine dehydratase. L-β-EHAsn is an atypical amino acid present in large amounts in urine but rarely detected in serum or most organs/tissues examined. Quantitative analyses of urinary L-β-EHAsn in young healthy volunteers revealed significant correlation between urinary L-β-EHAsn concentration and creatinine level. Further, for in-depth analyses of L-β-EHAsn, we developed a simple three-step synthetic method using trans-epoxysuccinic acid as the starting substance. In addition, our research revealed a strong inhibitory effect of l-b-EHAsn on mammalian serine racemase, responsible for producing D-serine, a co-agonist of the N-methyl-D-aspartate receptor involved in glutamatergic neurotransmission.

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Section: Enzyme Purificationmentioning

confidence: 99%

“…The initial velocity of L-serine dehydration catalyzed by mouse SR was monitored by a coupled assay with a lactic dehydrogenase (LDH) as described previously [25]. The LDH-coupled assay was carried out in a reaction mixture containing 50 mM…”

Section: Inhibition Assay Of Serine Racemasementioning

confidence: 99%

Urinary l-erythro-β-hydroxyasparagine—a novel serine racemase inhibitor and substrate of the Zn2+-dependent d-serine dehydratase

et al. 2021

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Molecular basis and functional development of enzymes related to amino acid metabolism

Yoshimura¹

2022

Bioscience, Biotechnology, and Biochemistry

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Enzymology, the study of enzyme structures and reaction mechanisms can be considered a classical discipline. However, enzymes cannot be freely designed to catalyze desired reactions yet, and enzymology is by no means a complete science. I have long studied the reaction mechanisms of enzymes related to amino acid metabolism, such as aminotransferases and racemases, which depend on pyridoxal 5'-phosphate, a coenzyme form of vitamin B6. During these studies, I have often been reminded that enzymatic reactions are extremely sophisticated processes based on chemical principles and enzyme structures, and have often been amazed at the evolutionary mechanisms that bestowed them with such structures. In this review, I described the reaction mechanism of various pyridoxal enzymes especially related to D-amino acids metabolism, whose roles in mammals have recently attracted attention. I hope to convey some of the significance and interest in enzymology through this review.

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Identification and characterization of a serine racemase in the silkworm Bombyx mori

Tanaka

Yoshimura

Hakamata

et al. 2022

The Journal of Biochemistry

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The pupae of lepidopterans contain high concentrations of endogenous d-serine. In the silkworm Bombyx mori, d-serine is negligible during the larval stage but increases markedly during the pupal stage, reaching 50% of the total free serine. However, the physiological function of d-serine and the enzyme responsible for its production are unknown. Herein, we identified a new type of pyridoxal 5′-phosphate (PLP)-dependent serine racemase (SR) that catalyzes the racemization of l-serine to d-serine in B. mori. This silkworm SR (BmSR) has an N-terminal PLP-binding domain that is homologous to mammalian SR and a C-terminal putative ligand-binding regulatory-like domain (ACT-like domain) that is absent in mammalian SR. Similar to mammalian SRs, BmSR catalyzes the racemization and dehydration of both serine isomers. However, BmSR is different from mammalian SRs as evidenced by its insensitivity to Mg2+/Ca2+ and Mg-ATP—which are required for activation of mammalian SRs—and high d-serine dehydration activity. At the pupal stage, the SR activity was predominantly detected in the fat body, which was consistent with the timing and localization of BmSR expression. The results are an important first step in elucidating the physiological significance of d-serine in lepidopterans.

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Mechanism of eukaryotic serine racemase-catalyzed serine dehydration

Cited by 3 publications

References 25 publications

Urinary l-erythro-β-hydroxyasparagine—a novel serine racemase inhibitor and substrate of the Zn2+-dependent d-serine dehydratase

Urinary l-erythro-β-hydroxyasparagine—a novel serine racemase inhibitor and substrate of the Zn2+-dependent d-serine dehydratase

Molecular basis and functional development of enzymes related to amino acid metabolism

Identification and characterization of a serine racemase in the silkworm Bombyx mori

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