title: Molecular heterogeneity of free PSA in sera of patients with benign and malignant prostate tumors.
abstract: OBJECTIVE
  To analyze free prostate-specific antigen (f-PSA) in sera from patients with prostate cancer (PCa) and benign prostatic hyperplasia (BPH), and to detect possible differences in subtypes as potential diagnostic parameters.
  
  
  METHODS
  PSA was purified from sera by an immunoaffinity procedure developed on the basis of oriented antibody immobilization, and subjected to size exclusion chromatography (SEC), Western blotting, and N-terminal amino acid sequencing.
  
  
  RESULTS
  The novel procedure allowed the purification of PSA with high yield from sera containing PSA <10 ng/ml. SEC under nonreducing conditions as well as Western blots demonstrated the presence of several molecular forms of f-PSA. Three of the smaller polypeptides exhibited the N-terminal sequence of PSA while one represented the C-terminal fragment Lys(146)-Pro(237). Shortening of some polypeptides by the N-terminal amino acid Ile(1) suggestive of aminopeptidase action was also observed. No propeptide sequence could be detected, and none of the bands from patient sera reacted with antibodies raised against propeptide antigens. BPH sera expressed higher proportions of smaller PSA fragments per unit p33, and contained significant amounts of fragments <14,000 which appeared to be very low or absent from most PCa sera.
  
  
  CONCLUSIONS
  f-PSA as obtained from BPH and PCa sera represents a heterogeneous fraction. The major component (p33) is not in the nicked form and does not contain proPSA. Diagnostic potential could arise from the quantitative differences of the smaller PSA derivatives seen between PCa and BPH sera.
authors.affiliation: Institut für Physiologische Chemie, Universitätskrankenhaus Eppendorf, Hamburg, Deutschland. hilz@uke-uni-hamburg.de
authors.forename: H
authors.initials: H
authors.lastname: Hilz
authors.forename: J
authors.initials: J
authors.lastname: Noldus
authors.forename: P
authors.initials: P
authors.lastname: Hammerer
authors.forename: F
authors.initials: F
authors.lastname: Buck
authors.forename: M
authors.initials: M
authors.lastname: Lück
authors.forename: H
authors.initials: H
authors.lastname: Huland
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D006801:Humans; D007164:Immunosorbents / Q000378:metabolism; D007527:Isoenzymes / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D017430:Prostate-Specific Antigen / Q000097:blood* / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011470:Prostatic Hyperplasia / Q000097:blood / Q000201:enzymology*; D011471:Prostatic Neoplasms / Q000097:blood / Q000201:enzymology*; D017421:Sequence Analysis
chemical_list: D007164:Immunosorbents; D007527:Isoenzymes; D010446:Peptide Fragments; D017430:Prostate-Specific Antigen