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Proteases with plasminogen activator activity in hamster oviduct
M Jiménez Díaz1, S Giunta, J Valz-Gianinet
1Instituto de Biología, Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Argentina.
Molecular Reproduction and Development
|December 22, 1999
Summary
Hamster oviductal fluid and tissue extracts exhibit esterase and amidase activity, primarily due to plasminogen activators like tissue-type plasminogen activator (tPA). Gonadotrophic hormones stimulate these enzyme secretions, suggesting a role in reproductive physiology.
Area of Science:
- Reproductive Biology
- Enzymology
- Biochemistry
Background:
- The physiological roles of most oviductal proteins remain largely uncharacterized.
- Oviductal secretions and tissues possess enzymatic activities that are not fully understood.
Purpose of the Study:
- To investigate the esterase and amidase activities in hamster oviductal secretions and tissue extracts.
- To characterize the properties of these enzymes and identify their potential identity as plasminogen activators.
- To explore the influence of gonadotrophic hormones on the synthesis and secretion of these enzymes.
Main Methods:
- Analysis of proteolytic-like esterase and amidase activity in oviductal tissue extracts (OE) and oviductal fluid (OF).
- Enzyme characterization using synthetic substrates (TAME, BAPNA), varying pH, and specific inhibitors (SBTI, TLCK, PMSF, benzamidine).
- Affinity chromatography using WGA-Sepharose to isolate glycoprotein fractions.
- Detection and quantification of plasminogen activators (PA) in tissue membrane fractions and oviductal secretions using plasminogen-dependent assays and specific inhibitors (amiloride).
Main Results:
- Oviductal secretions and tissue extracts displayed significant esterase and amidase activity, enhanced in superovulated hamsters, indicating hormonal stimulation.
- Enzymatic activity was calcium-dependent, optimal at pH 7.5, and inhibited by various protease inhibitors.
- Protease activity was identified as plasminogen activator (PA), with tissue-type plasminogen activator (tPA) being the predominant form (90%) in oviductal tissue membranes, while oviductal fluid showed limited plasminogen-dependent activity.
Conclusions:
- Hamster oviductal enzymes exhibit properties consistent with plasminogen activators, particularly tPA.
- Gonadotrophic hormones stimulate the synthesis and secretion of these proteolytic enzymes.
- The findings suggest a potential physiological role for these oviductal enzymes in reproductive processes.
