Detection and quantitation of glutamate carboxypeptidase II in human blood

0428605 - ÚOCHB 2015 RIV US eng J - Článek v odborném periodiku
Knedlík, Tomáš - Navrátil, Václav - Vik, V. - Pacík, D. - Šácha, Pavel - Konvalinka, Jan
Detection and quantitation of glutamate carboxypeptidase II in human blood.
Prostate. Roč. 74, č. 7 (2014), s. 768-780. ISSN 0270-4137. E-ISSN 1097-0045
Grant CEP: GA ČR GAP304/12/0847
Grant ostatní: OPPC(CZ) CZ.2.16/3.1.00/24016
Institucionální podpora: RVO:61388963
Klíčová slova: serum marker * glutamate carboxypeptidase II * plasma glutamate carboxypeptidase * prostate cancer * prostate-specific membrane antigen
Kód oboru RIV: CE - Biochemie
Impakt faktor: 3.565, rok: 2014

Glutamate carboxypeptidase II (GCPII) is a transmembrane enzyme that cleaves N-acetyl-L-aspartyl-L-glutamate (NAAG) in the brain. GCPII is highly expressed in the prostate and prostate cancer and might be associated with prostate cancer progression. Another exopeptidase, plasma glutamate carboxypeptidase (PGCP), was reported to be similar to GCPII and to share its NAAG-hydrolyzing activity. We performed a radioenzymatic assay with [H-3]NAAG as a substrate to detect and quantify the enzymatic activity of GCPII in plasma. Using a specific antibody raised against native GCPII (2G7), we immunoprecipitated GCPII from human plasma. We also cloned two PGCP constructs, expressed them in insect cells, and tested them for their NAAG-hydrolyzing activity. We detected GCPII protein in human plasma and found that its concentration ranges between 1.3 and 17.2 ng/ml in volunteers not diagnosed with prostate cancer. Recombinant PGCP was enzymatically active but exhibited no NAAG-hydrolyzing activity. GCPII is present in human blood, and its concentration within a healthy population varies. Recombinant PGCP does not hydrolyze NAAG, suggesting that GCPII alone is responsible for the NAAG-hydrolyzing activity observed in human blood. The potential correlation between GCPII serum levels and the disease status of prostate cancer patients will be further investigated.
Trvalý link: http://hdl.handle.net/11104/0233924