Expression and characterization of trypsinogen produced in the human male genital tract.
Trypsinogen is a serine proteinase produced mainly by the pancreas, but it has recently been found to be expressed also in several cancers such as ovarian and colon cancer and in vascular endothelial cells. In this study, we found that trypsinogen-1 and -2 are present at high concentrations (median levels, 0.4 and 0.5 mg/L, respectively) in human seminal fluid and purified them to homogeneity by immunoaffinity and anion exchange chromatography. Purified trypsinogen isoenzymes displayed a M(r) of 25 to 28 kd in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Most of the trypsinogen-1 purified from seminal fluid was enzymatically active whereas trypsinogen-2 occurred as the proform, which could be activated by enteropeptidase in vitro. Immunohistochemically, trypsinogen protein was detected in the human prostate, urethra, utriculus, ejaculatory duct, seminal vesicles, deferent duct, epididymal glands, and testis. Expression of trypsinogen mRNA in the same organs was demonstrated by in situ hybridization. Trypsinogen mRNA was also detected in the prostate and seminal vesicles by reverse transcriptase-polymerase chain reaction and Northern blotting. Isolated trypsin was shown to activate the proenzyme form of prostate-specific antigen. These results suggest that trypsinogen isoenzymes found in seminal fluid are produced locally in the male genital tract and that they may play a physiological role in the semen.
Paju
A
A
Departments of Clinical Chemistry and Pathology, Helsinki University Central Hospital, Helsinki, Finland. annukka.lukkonen@helsinki.fi
Bjartell
A
A
Zhang
W M
WM
Nordling
S
S
Borgström
A
A
Hansson
J
J
Stenman
U H
UH
D015152:Blotting, Northern; D005837:Genitalia, Male / Q000201:enzymology*; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D008297:Male; D017430:Prostate-Specific Antigen / Q000502:physiology; D012661:Semen / Q000201:enzymology; D014357:Trypsin / Q000494:pharmacology; D014362:Trypsinogen / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D015032:Zinc / Q000494:pharmacology
D007527:Isoenzymes; D014362:Trypsinogen; D014357:Trypsin; D017430:Prostate-Specific Antigen; D015032:Zinc