Molecular cloning of a peptidase against N-acetylaspartylglutamate from a rat hippocampal cDNA library

Tomasz Bzdega, Thomas Turi, Barbara Wroblewska, Dewei She, Hye Sun Chung, Hyun Kim, Joseph H. Neale

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

N-Acetylaspartylglutamate (NAAG) is the most prevalent peptide neurotransmitter in the mammalian nervous system. NAAG selectively activates the type 3 metabotropic glutamate receptor. It is inactivated by peptidase activity on the extracellular face of the plasma membrane of neurons and glia. The human gene that codes for prostate-specific membrane antigen (PSM) has been shown to produce peptidase activity against NAAG. We cloned the human PSM cDNA and used it to probe a rat hippocampal cDNA library. We identified a cDNA containing a complete coding region that possesses 83% homology with the PSM gene. The predicted 752-amino acid sequence has 85% identity and 91% similarity to the PSM sequence. CHO cells transfected with this cDNA expressed NAAG peptidase activity at a level similar to that obtained from rat brain membranes. The peptidase activity was inhibited by β-NAAG, quisqualate, and pteroylglutamate but not aspartylglutamate or pteroic acid. In situ hybridization data demonstrated the widespread distribution of the peptidase mRNA in the brain, consistent with the distribution of peptidase activity. The highest levers of hybridization were detected in the hippocampus, dentate gyrus, piriform cortex, choroid plexus of the ventricles, pineal gland, anterior pituitary, and supraoptic nucleus. Three transcripts (estimated at 5, 3.4, and 2.9 kb) were identified in northern blots of rat brain, while in rat kidney the third transcript appeared slightly smaller than 2.9 kb. With use of reverse transcriptase PCR with primers for the 5' end, the central region, and the 3' end of the hippocampal cDNA, the expected amplification products were obtained from rat brain RNA. Spinal cord yielded an amplification product only with primers for the 5' end of the hippocampal cDNA.

Original languageEnglish
Pages (from-to)2270-2277
Number of pages8
JournalJournal of Neurochemistry
Volume69
Issue number6
Publication statusPublished - 1997 Dec 1

Fingerprint

Glutamate Carboxypeptidase II
Cloning
Molecular Cloning
Gene Library
Rats
Peptide Hydrolases
Complementary DNA
Brain
aspartylglutamate
Amplification
Genes
Quisqualic Acid
Supraoptic Nucleus
Choroid Plexus
Pineal Gland
CHO Cells
RNA-Directed DNA Polymerase
Dentate Gyrus
Neurology
Cell membranes

Keywords

  • Cloning
  • Excitatory transmitter
  • Glutamate
  • N-Acetylaspartyl-grutamate
  • Peptidase

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Bzdega, T., Turi, T., Wroblewska, B., She, D., Chung, H. S., Kim, H., & Neale, J. H. (1997). Molecular cloning of a peptidase against N-acetylaspartylglutamate from a rat hippocampal cDNA library. Journal of Neurochemistry, 69(6), 2270-2277.

Molecular cloning of a peptidase against N-acetylaspartylglutamate from a rat hippocampal cDNA library. / Bzdega, Tomasz; Turi, Thomas; Wroblewska, Barbara; She, Dewei; Chung, Hye Sun; Kim, Hyun; Neale, Joseph H.

In: Journal of Neurochemistry, Vol. 69, No. 6, 01.12.1997, p. 2270-2277.

Research output: Contribution to journalArticle

Bzdega, T, Turi, T, Wroblewska, B, She, D, Chung, HS, Kim, H & Neale, JH 1997, 'Molecular cloning of a peptidase against N-acetylaspartylglutamate from a rat hippocampal cDNA library', Journal of Neurochemistry, vol. 69, no. 6, pp. 2270-2277.
Bzdega, Tomasz ; Turi, Thomas ; Wroblewska, Barbara ; She, Dewei ; Chung, Hye Sun ; Kim, Hyun ; Neale, Joseph H. / Molecular cloning of a peptidase against N-acetylaspartylglutamate from a rat hippocampal cDNA library. In: Journal of Neurochemistry. 1997 ; Vol. 69, No. 6. pp. 2270-2277.
@article{19ce1de3e2b649fe989a73ad93504806,
title = "Molecular cloning of a peptidase against N-acetylaspartylglutamate from a rat hippocampal cDNA library",
abstract = "N-Acetylaspartylglutamate (NAAG) is the most prevalent peptide neurotransmitter in the mammalian nervous system. NAAG selectively activates the type 3 metabotropic glutamate receptor. It is inactivated by peptidase activity on the extracellular face of the plasma membrane of neurons and glia. The human gene that codes for prostate-specific membrane antigen (PSM) has been shown to produce peptidase activity against NAAG. We cloned the human PSM cDNA and used it to probe a rat hippocampal cDNA library. We identified a cDNA containing a complete coding region that possesses 83{\%} homology with the PSM gene. The predicted 752-amino acid sequence has 85{\%} identity and 91{\%} similarity to the PSM sequence. CHO cells transfected with this cDNA expressed NAAG peptidase activity at a level similar to that obtained from rat brain membranes. The peptidase activity was inhibited by β-NAAG, quisqualate, and pteroylglutamate but not aspartylglutamate or pteroic acid. In situ hybridization data demonstrated the widespread distribution of the peptidase mRNA in the brain, consistent with the distribution of peptidase activity. The highest levers of hybridization were detected in the hippocampus, dentate gyrus, piriform cortex, choroid plexus of the ventricles, pineal gland, anterior pituitary, and supraoptic nucleus. Three transcripts (estimated at 5, 3.4, and 2.9 kb) were identified in northern blots of rat brain, while in rat kidney the third transcript appeared slightly smaller than 2.9 kb. With use of reverse transcriptase PCR with primers for the 5' end, the central region, and the 3' end of the hippocampal cDNA, the expected amplification products were obtained from rat brain RNA. Spinal cord yielded an amplification product only with primers for the 5' end of the hippocampal cDNA.",
keywords = "Cloning, Excitatory transmitter, Glutamate, N-Acetylaspartyl-grutamate, Peptidase",
author = "Tomasz Bzdega and Thomas Turi and Barbara Wroblewska and Dewei She and Chung, {Hye Sun} and Hyun Kim and Neale, {Joseph H.}",
year = "1997",
month = "12",
day = "1",
language = "English",
volume = "69",
pages = "2270--2277",
journal = "Journal of Neurochemistry",
issn = "0022-3042",
publisher = "Wiley-Blackwell",
number = "6",

}

TY - JOUR

T1 - Molecular cloning of a peptidase against N-acetylaspartylglutamate from a rat hippocampal cDNA library

AU - Bzdega, Tomasz

AU - Turi, Thomas

AU - Wroblewska, Barbara

AU - She, Dewei

AU - Chung, Hye Sun

AU - Kim, Hyun

AU - Neale, Joseph H.

PY - 1997/12/1

Y1 - 1997/12/1

N2 - N-Acetylaspartylglutamate (NAAG) is the most prevalent peptide neurotransmitter in the mammalian nervous system. NAAG selectively activates the type 3 metabotropic glutamate receptor. It is inactivated by peptidase activity on the extracellular face of the plasma membrane of neurons and glia. The human gene that codes for prostate-specific membrane antigen (PSM) has been shown to produce peptidase activity against NAAG. We cloned the human PSM cDNA and used it to probe a rat hippocampal cDNA library. We identified a cDNA containing a complete coding region that possesses 83% homology with the PSM gene. The predicted 752-amino acid sequence has 85% identity and 91% similarity to the PSM sequence. CHO cells transfected with this cDNA expressed NAAG peptidase activity at a level similar to that obtained from rat brain membranes. The peptidase activity was inhibited by β-NAAG, quisqualate, and pteroylglutamate but not aspartylglutamate or pteroic acid. In situ hybridization data demonstrated the widespread distribution of the peptidase mRNA in the brain, consistent with the distribution of peptidase activity. The highest levers of hybridization were detected in the hippocampus, dentate gyrus, piriform cortex, choroid plexus of the ventricles, pineal gland, anterior pituitary, and supraoptic nucleus. Three transcripts (estimated at 5, 3.4, and 2.9 kb) were identified in northern blots of rat brain, while in rat kidney the third transcript appeared slightly smaller than 2.9 kb. With use of reverse transcriptase PCR with primers for the 5' end, the central region, and the 3' end of the hippocampal cDNA, the expected amplification products were obtained from rat brain RNA. Spinal cord yielded an amplification product only with primers for the 5' end of the hippocampal cDNA.

AB - N-Acetylaspartylglutamate (NAAG) is the most prevalent peptide neurotransmitter in the mammalian nervous system. NAAG selectively activates the type 3 metabotropic glutamate receptor. It is inactivated by peptidase activity on the extracellular face of the plasma membrane of neurons and glia. The human gene that codes for prostate-specific membrane antigen (PSM) has been shown to produce peptidase activity against NAAG. We cloned the human PSM cDNA and used it to probe a rat hippocampal cDNA library. We identified a cDNA containing a complete coding region that possesses 83% homology with the PSM gene. The predicted 752-amino acid sequence has 85% identity and 91% similarity to the PSM sequence. CHO cells transfected with this cDNA expressed NAAG peptidase activity at a level similar to that obtained from rat brain membranes. The peptidase activity was inhibited by β-NAAG, quisqualate, and pteroylglutamate but not aspartylglutamate or pteroic acid. In situ hybridization data demonstrated the widespread distribution of the peptidase mRNA in the brain, consistent with the distribution of peptidase activity. The highest levers of hybridization were detected in the hippocampus, dentate gyrus, piriform cortex, choroid plexus of the ventricles, pineal gland, anterior pituitary, and supraoptic nucleus. Three transcripts (estimated at 5, 3.4, and 2.9 kb) were identified in northern blots of rat brain, while in rat kidney the third transcript appeared slightly smaller than 2.9 kb. With use of reverse transcriptase PCR with primers for the 5' end, the central region, and the 3' end of the hippocampal cDNA, the expected amplification products were obtained from rat brain RNA. Spinal cord yielded an amplification product only with primers for the 5' end of the hippocampal cDNA.

KW - Cloning

KW - Excitatory transmitter

KW - Glutamate

KW - N-Acetylaspartyl-grutamate

KW - Peptidase

UR - http://www.scopus.com/inward/record.url?scp=0030658978&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030658978&partnerID=8YFLogxK

M3 - Article

C2 - 9375657

AN - SCOPUS:0030658978

VL - 69

SP - 2270

EP - 2277

JO - Journal of Neurochemistry

JF - Journal of Neurochemistry

SN - 0022-3042

IS - 6

ER -