Expression and Characterization of Trypsinogen Produced in the Human Male Genital Tract

Abstract

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.

Expression of Trypsinogen-1, Trypsinogen-2, and Tumor-Associated Trypsin Inhibitor in Ovarian Cancer

“…Water was used as a negative control and cDNA from COLO 205 colon adenocarcinoma cells as a positive control in all experiments. The agarose gel electrophoresis and sequencing of the PCR products were carried out as described previously (40).…”

Prostatic trypsin-like kallikrein-related peptidases (KLKs) and other prostate-expressed tryptic proteinases as regulators of signalling via proteinase-activated receptors (PARs)

“…These include secreted enzymes, such as trypsin I, II (Paju et al, 2000;Bjartell et al, 2005) and IV (Takeuchi et al, 1999;Cottrell et al, 2004); other more complex secreted serine proteinases, containing modular nonproteolytic domains, such as factor XIIa (Takeuchi et al, 1999), plasma kallikrein (Neth et al, 2001;Fink et al, 2007), urokinase (also known as urinary plasminogen activator, uPA; Gavrilov et al, 2001;Riddick et al, 2005;Usher et al, 2005) and protein C (He et al, 1995;Takeuchi et al, 1999); the multi-serine proteinase domain containing enzymes polyserase-2 and -3 (Cal et al, 2005(Cal et al, , 2006; the glycosyl-phosphatidylinositol membraneanchored serine proteinase prostasin (Chen et al, 2001); as well as nine members of the type II transmembrane serine proteinase (TTSP) sub-family (Hooper et al, 2001;Netzel-Arnett et al, 2003) including hepsin, enteropeptidase (Cottrell et al, 2004), matriptase/MT-SP1 (Saleem et al, 2006), TMPRSS2 (Lin et al, 1999), TMPRSS3 (Scott et al, 2001), TMPRSS13 (Kim et al, 2001), human airway trypsin-like proteinase (HAT) (Hahner et al, 2005), DESC1 (Lang and Schuller, 2001) and the multi-catalytic domain TTSP polyserase-1 (Okumura et al, 2006). It is also interesting to note the co-expression in prostatederived cell lines of trypsinogen I, II and IV and the in vivo Brought to you by | University of Queensland -UQ Library Authenticated Download Date | 9/14/15 3:38 AM activator of these enzymes, the TTSP enteropeptidase (Light and Janska, 1989;Kitamoto et al, 1994).…”

Editorial Comment on: Increased Expression of Tumor-Associated Trypsin Inhibitor, TATI, in Prostate Cancer and in Androgen-Independent 22Rv1 Cells

“…Tumours require protelolytic activity for invasion, and trypsin is a potent activator of MMPs associated with tissue remodelling and invasion, it is therefore not completely surprising that various tumours e.g. ovarian [4], lung [5] and gastric [6] are reported to express trypsinogens and TATI [7]. More recently however it has been reported that trypsinogens are expressed in human seminal fluid suggesting one possible source could be the prostate gland [7].…”

“…ovarian [4], lung [5] and gastric [6] are reported to express trypsinogens and TATI [7]. More recently however it has been reported that trypsinogens are expressed in human seminal fluid suggesting one possible source could be the prostate gland [7]. Trypsin has been demonstrated to be expressed in the human prostate gland and indeed be involved both with activation of proPSA and degradation of PSA in the seminal plasma [7].…”