title: Rapid determination of free D-serine with chicken D-serine dehydratase.
abstract: We have developed a simple, rapid, and inexpensive method of measuring the concentration of intrinsic free D-serine in tissue samples. This method uses chicken D-serine dehydratase in an enzymatic reaction to produce pyruvate, which is detected spectrophotometrically. Pyridoxal 5'-phosphate (PLP), a cofactor of D-serine dehydratase, increased pyruvate formation by 28%. The presence of Zn(2+) or ethylenediaminetetraacetic acid (EDTA) did not have any effect on pyruvate formation under the present assay conditions. In addition, this method was not affected by the presence of a large excess of L-serine, nor by the presence of tissue extracts, and accurately determined concentrations of 2-30 μM (200 pmol-3 nmol) of D-serine. The entire assay requires only 60 min. With this method, we determined the concentration of D-serine in various silkworm tissues. The results were in agreement with high performance liquid chromatography measurements. We found high concentrations of D-serine in silkworm larvae at day 3 of the fifth instar; specifically, 509 nmol g(-1) wet tissue in the midgut, 434 nmol g(-1) in the ovary, and 353 nmol g(-1) in the testis.
authors.affiliation: Department of Materials and Applied Chemistry, College of Science and Technology, Nihon University, Kanda-Surugadai, Tokyo 101-8308, Japan.
authors.forename: Chihiro
authors.initials: C
authors.lastname: Suzuki
authors.forename: Masahito
authors.initials: M
authors.lastname: Murakami
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Yokobori
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tanaka
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Ishida
authors.forename: Kihachiro
authors.initials: K
authors.lastname: Horiike
authors.forename: Yoko
authors.initials: Y
authors.lastname: Nagata
mesh_terms: D000818:Animals; D012831:Bombyx; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D003124:Colorimetry / Q000379:methods*; D057075:Enzyme Assays; D006836:Hydro-Lyases / Q000737:chemistry / Q000378:metabolism*; D007814:Larva / Q000737:chemistry / Q000254:growth & development / Q000378:metabolism; D016014:Linear Models; D009928:Organ Specificity; D011732:Pyridoxal Phosphate / Q000378:metabolism; D019289:Pyruvic Acid / Q000737:chemistry / Q000378:metabolism; D012694:Serine / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism
chemical_list: D012694:Serine; D011732:Pyridoxal Phosphate; D019289:Pyruvic Acid; D006836:Hydro-Lyases; C101950:D-serine dehydratase