Recombinant glutamate carboxypeptidase II (prostate specific membrane antigen--PSMA)--cellular localization and bioactivity analyses.
Glutamate Carboxypeptidase II (also known as Prostate Specific Membrane Antigen-PSMA) is an important marker in the diagnosis of prostate cancer, however, relatively little is known about its biochemical and structure-function characteristics. We have expressed mutant forms of PSMA and have started to address the roles of three putative domains of PSMA in its cellular localization and peptidase activity. Three mutants, a full-length recombinant PSMA (rPSMA-FL), one expressing only the proposed extracellular domain of PSMA (rPSMA-ECD) and one form omitting the proposed transmembrane domain (rPSMA-deltaTMD) have been produced in human cells via a mammalian expression vector system. We show that rPSMA-FL is associated with the cell surface membrane; so too is rPSMA-deltaTMD even though it lacks the proposed transmembrane domain, whereas rPSMA-ECD has a cytosolic localization. Only rPSMA-FL retains functional hydrolytic activity and is similarly glycosylated to PSMA found in the cultured prostate cancer cell line LNCaP.
D000954:Antigens, Surface / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics; D043425:Glutamate Carboxypeptidase II / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection; D014316:Tritium
D000954:Antigens, Surface; D011994:Recombinant Proteins; D014316:Tritium; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II