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Trypsin‐like proteolytic contamination of commercially available psa purified from human seminal fluid
Author(s) -
Manning Michael L.,
Kostova Maya,
Williams Simon A.,
Denmeade Samuel R.
Publication year - 2011
Publication title -
the prostate
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.295
H-Index - 123
eISSN - 1097-0045
pISSN - 0270-4137
DOI - 10.1002/pros.22474
Subject(s) - prostate specific antigen , trypsin , prostate cancer , proteases , aprotinin , proteolysis , protease , prostate , serine protease , in vitro , antigen , lncap , chemistry , cancer , microbiology and biotechnology , medicine , biology , biochemistry , immunology , enzyme
BACKGROUND Prostate‐Specific Antigen (PSA) is a serine protease whose expression is maintained in all stages of prostate cancer. A role for PSA in the pathobiology for prostate cancer has not been firmly established. Experimental studies to date support a role for PSA through mechanisms such as release or processing of growth factors and degradation of the extracellular matrix. Exposure of prostate cancer cells to exogenous PSA also results in gene expression changes. These in vitro and biochemical assays rely on the use of commercially available PSA. Contamination of these commercial preparations can significantly impact the results of these in vitro studies. METHODS We characterized PSA and trypsin‐like activity of PSA preparations obtained from three commercial sources: Calbiochem, Fitzgerald, and AbD Serotec. Silver stained gels were used to compare the purity of each preparation and mass spectrometry was performed to characterize contaminating proteases. RESULTS PSA activity varied between PSA preparations with AbD Serotec PSA having highest degree of activity. Significant trypsin‐like activity, which was inhibited by aprotinin, was observed in PSA preparations from Calbiochem and Fitzgerald, but not AbD Serotec. These former two PSA preparations also contained the greatest degree of non‐PSA contaminants by silver stain and mass spectrometry. CONCLUSIONS Commercially available preparations of PSA contain contaminating proteins, including trypsin‐like protease activity, that could potentially complicate the interpretation of results obtained from in vitro studies assessing PSA proteolysis of potential protein substrates and effects of PSA on gene expression. Prostate 72:1233–1238, 2012. © 2011 Wiley Periodicals, Inc.