Cookies on this website
We use cookies to ensure that we give you the best experience on our website. If you click 'Continue' we'll assume that you are happy to receive all cookies and you won't see this message again. Click 'Find out more' for information on how to change your cookie settings.

Measurements of the prostate-specific antigen (PSA) levels in blood are widely used as diagnostic, predictive and prognostic marker of prostate disease. The selective detection of molecular forms of PSA can contribute clinically to meaningful enhancements of the conventional PSA-test. As it is plausible that an in-depth search for structural variants of PSA gene products may increase our ability to discriminate distinct patho-biological basis and stages of prostate diseases, we have developed a multi-step protocol comprising gel-based methods followed by mass spectrometric identification. Our current aim was to provide a comprehensive identification of PSA variants occurring in seminal fluid. We provide a proof-of-principle for this multiple step analytical approach to identify multiple PSA variants from complex biological samples that revealed distinct molecular characteristics. In addition, sequence-annotated protein bands in SDS-PAGE gels were compared to those detected by Western blots, and by monitoring the enzymatic activity in zymogram gels, using gelatin as a substrate. The high accuracy annotations were obtained by fast turnaround MALDI-Orbitrap analysis from excised and digested gel bands. Multiple PSA forms were identified utilizing a combination of MASCOT and SEQUEST search engines.

Original publication

DOI

10.1016/j.jprot.2010.01.008

Type

Journal article

Journal

J Proteomics

Publication Date

18/04/2010

Volume

73

Pages

1137 - 1147

Keywords

Amino Acid Sequence, Densitometry, Electrophoresis, Polyacrylamide Gel, Humans, Male, Mass Spectrometry, Molecular Sequence Data, Peptides, Prostate, Prostate-Specific Antigen, Proteomics, Reproducibility of Results, Semen, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization