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. Copyright (C) 1998 Federation of European Biochemical Societies.
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