Isolation and expression of a rat brain cDNA encoding glutamate carboxypeptidase II.
N-acetylated alpha-linked acidic dipeptidase (NAALADase) hydrolyzes acidic peptides, such as the abundant neuropeptide N-acetyl-alpha-L-aspartyl-L-glutamate (NAAG), thereby generating glutamate. Previous cDNA cloning efforts have identified a candidate rat brain NAALADase partial cDNA, and Northern analyses have identified a family of related RNA species that are found only in brain and other NAALADase-expressing cells. In this report, we describe the cloning of a set of rat brain cDNAs that describe a full-length NAALADase mRNA. Transient transfection of a full-length cDNA into the PC3 cell line confers NAAG-hydrolyzing activity that is sensitive to the NAALADase inhibitors quisqualic acid and 2-(phosphonomethyl)glutaric acid. Northern hybridization detects the expression of three similar brain RNAs approximately 3,900, 3,000, and 2,800 nucleotides in length. In situ hybridization histochemistry shows that NAALADase-related mRNAs have an uneven regional distribution in rat brain and are expressed predominantly by astrocytes as demonstrated by their colocalization with the astrocyte-specific marker glial fibrillary acidic protein.
Luthi-Carter
R
R
Department of Psychiatry, Massachusetts General Hospital-East, Charlestown, MA 02129, USA.
Berger
U V
UV
Barczak
A K
AK
Enna
M
M
Coyle
J T
JT
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology*; D002268:Carboxypeptidases / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004151:Dipeptides / Q000378:metabolism; D043425:Glutamate Carboxypeptidase II; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009479:Neuropeptides / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins; D017422:Sequence Analysis, DNA; D014018:Tissue Distribution
D018076:DNA, Complementary; D004151:Dipeptides; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D011994:Recombinant Proteins; C027172:isospaglumic acid; D002268:Carboxypeptidases; D043425:Glutamate Carboxypeptidase II