Characterization and localization of a human serine racemase.

D-serine is present in the mammalian central nervous system, where it acts as one of the co-activators of N-methyl-D aspartate receptors. Synthesis of D-serine is catalyzed by the serine racemase enzyme. The current studies report on the isolation of a cDNA encoding a human serine racemase (SRR) from the human neuronal like cell line, NT2N. The SRR gene was localized on chromosome 17q13. The full-length cDNA has 1020 nucleotides which encode for a protein of 340 amino acids. The human protein shares 89% sequence identity with the mouse serine racemase. Human embryonic kidney 293 cells transiently transfected with this SRR gene were able to produce d-serine, indicating that the sequence encodes for an active enzyme. In Northern blot analysis the SRR mRNA was expressed in human brain, heart, skeletal muscle, kidney and liver tissues. Different splice forms of SRR were present in the peripheral tissues. Transcripts of at least three different sizes were present in heart and kidney, while in Western blot analysis multiple bands of different sizes were observed. Immunohistochemical studies, using a polyclonal anti-human serine racemase antibody, revealed a peripheral expression of serine racemase protein in human cardiac myocytes and convoluted tubules of the kidney. Experiments in non-human primate brain demonstrated the localization of SRR in amygdala nuclei, cortex, thalamus and hippocampus. Co-localization studies in the hippocampus demonstrated the exclusive expression of serine racemase in glial cells. The cloning of a functional human serine racemase and its expression in central nervous system of primates support a role for D-serine in neuronal activity. Furthermore, its presence in human periphery such as in heart and kidney suggest a potential biological role for D-serine in the regulation of N-methyl-D-aspartate (NMDA) receptor activity in these peripheral organs as well.