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Recombinant Glutamate Carboxypeptidase II (Prostate Specific Membrane Antigen—PSMA)—Cellular Localization and Bioactivity Analyses

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Abstract

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-ΔTMD) 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-ΔTMD 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.

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Authors and Affiliations

  1. Department of Biochemistry, University of Missouri, Columbia, Missouri

    Mark A. Meighan, Marie T. Dickerson & Susan L. Deutscher

  2. Harry S. Truman Memorial Veteran's Hospital, Columbia, Missouri

    Olga Glinskii & Vladislav V. Glinsky

  3. Department of Microbiology and Molecular Cell Biology and Virginia Prostate Center, Eastern Virginia Medical School, Norfolk, Virginia

    George L. Wright Jr.

Authors
  1. Mark A. Meighan
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  2. Marie T. Dickerson
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  3. Olga Glinskii
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  4. Vladislav V. Glinsky
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  5. George L. Wright Jr.
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  6. Susan L. Deutscher
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Correspondence to Susan L. Deutscher.

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Meighan, M.A., Dickerson, M.T., Glinskii, O. et al. Recombinant Glutamate Carboxypeptidase II (Prostate Specific Membrane Antigen—PSMA)—Cellular Localization and Bioactivity Analyses. J Protein Chem 22, 317–326 (2003). https://doi.org/10.1023/A:1025381921943

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