Enzymes involved in the metabolism of gamma-hydroxybutyrate in SH-SY5Y cells: identification of an iron-dependent alcohol dehydrogenase ADHFe1.
The metabolism of the endogenous metabolite gamma-hydroxybutyrate (GHB) has been studied in a human neuroblastoma cell line SH-SY5Y as a model for examining neuronal metabolism. We show that GHB can be synthesized and released from these cells, indicating that pathways for GHB synthesis and secretion are present. Activities for the major enzymes that are involved in GHB metabolism are reported, and transcripts for AKR1A1, AKR7A2, ALDH5A1 and GABA-T can be detected by RT-PCR. We also demonstrate the presence of the ADHFe1 transcript, a gene that has been reported to encode a hydroxyacid-oxoacid transhydrogenase (HOT). We show that the ADHFe1 gene is related to bacterial GHB dehydrogenases and has a conserved NAD-binding site. The potential for using the SH-SY5Y cell line for investigating GHB catabolism is discussed.
Lyon
Robert C
RC
Strathclyde Institute of Pharmacy & Biomedical Sciences, University of Strathclyde, Glasgow, Scotland, UK.
Johnston
Stuart M
SM
Panopoulos
Andreas
A
Alzeer
Samar
S
McGarvie
Gail
G
Ellis
Elizabeth M
EM
D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D045744:Cell Line, Tumor; D017931:DNA Primers; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007501:Iron / Q000378:metabolism*; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D012978:Sodium Oxybate / Q000378:metabolism*
D017931:DNA Primers; D012978:Sodium Oxybate; D007501:Iron; D000426:Alcohol Dehydrogenase