title: Biological activity of prostate-specific antigen isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroelution.
abstract: Human prostate-specific antigen (PSA), a 33 kDa kallikrein-like serine protease, occurring in the prostate, in seminal plasma and in blood, was prepared under nonreducing conditions in an enzymatically active form from seminal plasma by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by fast copper staining, electroelution from gel slices and dialysis against isotonic phosphate-buffered saline (PBS). Enzymatic activity was demonstrated for the first time directly by cleavage of semenogelin, one of the biological substrates of PSA, isolated by the same procedure, i.e. SDS-PAGE and electroelution, but from seminal vesicle fluid. The purified PSA formed SDS-stable complexes with the two major extracellular protease inhibitors in blood, alpha 1-antichymotrypsin (alpha 1-ACH) and alpha 2-macroglobulin (alpha 2-M). PSA isolated under reducing conditions was enzymatically inactive and could not bind to the protease inhibitors alpha 1-ACH and alpha 2-M.
authors.affiliation: Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie, Universität Hamburg, Germany.
authors.forename: U
authors.initials: U
authors.lastname: Tessmer
authors.forename: T
authors.initials: T
authors.lastname: Quack
authors.forename: F
authors.initials: F
authors.lastname: Donn
authors.forename: A
authors.initials: A
authors.lastname: Leuner
authors.forename: R
authors.initials: R
authors.lastname: Dernick
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D012739:Gonadal Steroid Hormones / Q000378:metabolism; D006801:Humans; D008297:Male; D017430:Prostate-Specific Antigen / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012661:Semen / Q000201:enzymology*; D029607:Seminal Vesicle Secretory Proteins; D000514:alpha 1-Antichymotrypsin / Q000737:chemistry; D000511:alpha-Macroglobulins / Q000737:chemistry
chemical_list: D012739:Gonadal Steroid Hormones; D029607:Seminal Vesicle Secretory Proteins; D000514:alpha 1-Antichymotrypsin; D000511:alpha-Macroglobulins; C424482:seminal vesicle-specific antigen; D017430:Prostate-Specific Antigen