Two human glutamate decarboxylases, 65-kDa GAD and 67-kDa GAD, are each encoded by a single gene.
We report the isolation and sequencing of cDNAs encoding two human glutamate decarboxylases (GADs; L-glutamate 1-carboxy-lyase, EC 4.1.1.15), GAD65 and GAD67. Human GAD65 cDNA encodes a Mr 65,000 polypeptide, with 585 amino acid residues, whereas human GAD67 encodes a Mr 67,000 polypeptide, with 594 amino acid residues. Both cDNAs direct the synthesis of enzymatically active GADs in bacterial expression systems. Each cDNA hybridizes to a single species of brain mRNA and to a specific set of restriction fragments in human genomic DNA. In situ hybridization of fluorescently labeled GAD probes to human chromosomes localizes the human GAD65 gene to chromosome 10p11.23 and the human GAD67 gene to chromosome 2q31. We conclude that GAD65 and GAD67 each derive from a single separate gene. The cDNAs we describe should allow the bacterial production of test antigens for the diagnosis and prediction of insulin-dependent diabetes mellitus.
Bu
D F
DF
Department of Biology, University of California, Los Angeles 90024.
Erlander
M G
MG
Hitz
B C
BC
Tillakaratne
N J
NJ
Kaufman
D L
DL
Wagner-McPherson
C B
CB
Evans
G A
GA
Tobin
A J
AJ
D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D001921:Brain / Q000201:enzymology; D002874:Chromosome Mapping; D002879:Chromosomes, Human, Pair 10; D002889:Chromosomes, Human, Pair 2; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D015342:DNA Probes; D003922:Diabetes Mellitus, Type 1 / Q000175:diagnosis / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005333:Fetus; D015723:Gene Library; D005796:Genes; D005968:Glutamate Decarboxylase / Q000235:genetics*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D015342:DNA Probes; D012333:RNA, Messenger; D004247:DNA; D005968:Glutamate Decarboxylase