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Biosynthesis of d ‐aspartate in mammalian cells 1
Author(s) -
Long Zhiqun,
Homma Hiroshi,
Lee Jen-Ai,
Fukushima Takeshi,
Santa Tomofumi,
Iwatsubo Takeshi,
Yamada Ryo-hei,
Imai Kazuhiro
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00986-7
Subject(s) - biosynthesis , chemistry , biochemistry , microbiology and biotechnology , biology , gene
In this communication, we demonstrate that d ‐aspartate ( d ‐Asp) is synthesized in pheochromocytoma cells (PC12). To our knowledge this is the first report of biosynthesis of d ‐Asp in mammalian cells. Synthesis of d ‐Asp was demonstrated by its time‐dependent accumulation in the cell culture, and by the fact that this accumulation was proportional to the number of inoculated cells. d ‐Asp in PC12 cells was identified by (i) co‐elution with authentic d ‐Asp on two different HPLC columns, an octadesyl silica column and a Pirkle‐type chiral column, (ii) reversed elution order of d ‐Asp and l ‐Asp on another Pirkle‐type chiral column with an opposite configuration, and (iii) sensitivity to d ‐Asp oxidase. In the cells the amount of d ‐Asp was approx. 12–14% of total Asp and no other investigated d ‐amino acid was detected. The amount of d ‐Asp did not increase during the culture of mouse 3T3 fibroblasts and human neuroblastoma NB‐1 cells. Immunocytochemical staining with anti‐ d ‐Asp antiserum demonstrated that d ‐Asp synthesized is present in the cytoplasm of the cells.