PMID: 11905994Mar 22, 2002Paper

Substrate specificity, inhibition and enzymological analysis of recombinant human glutamate carboxypeptidase II

Journal of Neurochemistry
Cyril BarinkaJan Konvalinka

Abstract

Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a membrane peptidase expressed in a number of tissues such as kidney, prostate and brain. The brain form of GCPII (also known as NAALADase) cleaves N-acetyl-aspartyl glutamate to yield free glutamate. Animal model experiments show that inhibition of GCPII prevents neuronal cell death during experimental ischaemia. GCPII thus represents an important target for the treatment of neuronal damage caused by excess glutamate. In this paper we report expression of an extracellular portion of human glutamate carboxypeptidase II (amino acids 44-750) in Drosophila Schneider's cells and its purification to homogeneity. A novel assay for hydrolytic activity of recombinant human GCPII (rhGCPII), based on fluorimetric detection of released alpha-amino groups was established, and used for its enzymological characterization. rhGCPII does not show dipeptidylpeptidase IV-like activity assigned to the native form of the enzyme previously. Using a complete set of protected dipeptides, substrate specificity of rhGCPII was elucidated. In addition to the previously described substrates, four novel compounds, Ac-Glu-Met, Ac-Asp-Met and, surprisingly, Ac-Ala-Glu and Ac-Ala-Met were identified as sub...Continue Reading

Citations

Nov 5, 2002·Analytical Biochemistry·Isabelle KamgaClifford E Berkman
Jul 20, 2002·Bioorganic & Medicinal Chemistry Letters·Takashi TsukamotoJames K Coward
Jun 12, 2003·Bioorganic & Medicinal Chemistry Letters·Doris StoermerTakashi Tsukamoto
Dec 23, 2003·The International Journal of Biochemistry & Cell Biology·Petr BusekAleksi Sedo
Apr 26, 2003·Current Opinion in Chemical Biology·Richard C HolzSabina I Swierczek
Aug 28, 2003·Current Opinion in Chemical Biology·Jonathan S Rosenblum, John W Kozarich
Sep 20, 2011·Journal of Medicinal Chemistry·Anna PlechanovováCyril Bařinka
Mar 3, 2007·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Cyril BarinkaJacek Lubkowski
Oct 18, 2012·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Shahriar KoochekpourRobert L Vessella
Nov 17, 2009·Journal of Nuclear Medicine : Official Publication, Society of Nuclear Medicine·Suzanne E LapiElla F Jones
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Jul 31, 2013·Biochemical and Biophysical Research Communications·Suk Kyung LeeSang Ick Park
Jan 24, 2016·Protein Engineering, Design & Selection : PEDS·Cyril BarinkaArne Skerra
Jun 1, 2008·Expert Opinion on Medical Diagnostics·Aleksi SedoJonathan S Duke-Cohan
Feb 1, 2008·Journal of Molecular Biology·Cyril BarinkaJacek Lubkowski
Sep 4, 2007·Bioorganic & Medicinal Chemistry·Marc O AndersonClifford E Berkman
Sep 11, 2007·Drug Discovery Today·Takashi TsukamotoBarbara S Slusher
Dec 8, 2006·Neuroscience·P SáchaJ Konvalinka
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Aug 24, 2007·The FEBS Journal·Petra MlcochováJan Konvalinka
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