| Thiophilic-interaction chromatography of enzymatically active tissue prostate-specific antigen (T-PSA) and its modulation by zinc ions. | |
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MedLine Citation:
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PMID: 18083072 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Prostate-specific antigen (PSA) is a serine protease secreted both by normal prostate glandular epithelial cells and prostate cancer cells. We explored "thiophilic-interaction chromatography" (TIC) to isolate tissue prostate-specific antigen (T-PSA) from fresh human prostate cancer tissue harvested by radical prostatectomy for the purpose to characterize T-PSA for its enzymatic activity and sensitivity to zinc ions. We have shown, for the first time, that T-PSA has strong affinity for the thiophilic gel (T-gel). The average recovery of T-PSA from T-gel is over 87%. The presence of PSA in the column eluate was confirmed by ELISA and SDS/PAGE. Western blot developed with monoclonal antibody to PSA revealed that T-PSA was predominantly in the "free" form having a molecular weight of 33 kDa. Furthermore, T-PSA was found to be enzymatically active. T-PSA was found to be less enzymatically active as compared to seminal plasma PSA. The inhibition of enzymatic activity of both f-PSA and T-PSA over a wide range of concentrations of Zn(2+) ions (10nM to 50 microM) was comparable. In contrast, the enzymatic activity of chymotrypsin, another serine-protease, was affected differently. At higher concentrations of Zn(2+) (10 microM and higher) the enzymatic activity of chymotrypsin was inhibited, whereas, at lower concentrations of Zn(2+) (5 microM and lower), the enzymatic activity was enhanced. |
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Authors:
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A K Satheesh Babu; M A Vijayalakshmi; Gary J Smith; Kailash C Chadha |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2007-12-04 |
Journal Detail:
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Title: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Volume: 861 ISSN: 1570-0232 ISO Abbreviation: J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. Publication Date: 2008 Jan |
Date Detail:
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Created Date: 2008-01-28 Completed Date: 2008-03-17 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 101139554 Medline TA: J Chromatogr B Analyt Technol Biomed Life Sci Country: Netherlands |
Other Details:
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Languages: eng Pagination: 227-35 Citation Subset: IM |
Affiliation:
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Centre for Bio-Separation Technology, VIT University, Vellore, India. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Blotting, Western Catalysis / drug effects Chromatography, Affinity Chymotrypsin / metabolism Dose-Response Relationship, Drug Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay Humans Male Prostate-Specific Antigen / antagonists & inhibitors, isolation & purification, metabolism* Prostatic Neoplasms / metabolism*, pathology Substrate Specificity Zinc / pharmacology* |
| Grant Support | |
ID/Acronym/Agency:
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R21 CA113950-02/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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7440-66-6/Zinc; EC 3.4.21.1/Chymotrypsin; EC 3.4.21.77/Prostate-Specific Antigen |
| Comments/Corrections | |
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