Proteases with plasminogen activator activity in hamster oviduct.
At present the physiological role of most oviductal proteins remains unknown. In this work, we present evidence that the oviductal secretion as well as the crude oviductal tissue-extract show proteolytic-like esterase and amidase activity. The proteolytic activity of the oviductal enzymes was higher in the oviducts of superovulated hamster females than in those of normal ones, indicating that gonadotrophic hormones would stimulate the synthesis and secretion of these enzymes. Some of their properties were analyzed in the 15,600-g supernatant of both oviductal tissue extracts (OE) and oviductal fluid (OF). The enzymatic activity toward the synthetic substrates p-tosyl-l-arginine methyl ester-HCl (TAME) and alpha-N-benzoyl-dl-arginine-p-nitroanilide HCl (BAPNA) was activated by calcium ions, reached a maximum at pH 7.5, and was inhibited by soybean trypsin inhibitor (SBTI), N-alpha-p-tosyl-l-lysine chloromethyl ketone HCl (TLCK), phenyl methyl sulfonyl fluoride (PMSF), and benzamidine. The OE glycoprotein fraction recognized by WGA-Sepharose affinity columns (37% total proteins) showed proteolytic activity with properties similar to the OE and OF enzymes. The protease activity could be ascribed to a plasminogen activator (PA) detected in the Triton X-100 treated tissue crude membrane fraction (Triton-CMF) and in the oviductal secretion of the superovulated females. In the Triton-CMF fraction, 100% of the proteolytic activity was plasminogen-dependent. The use of amiloride, a selective urokinase-type plasminogen activator (uPA) inhibitor, shows that 90% of this activity was due to a tissue-type plasminogen activator (tPA) and 10% to uPA whereas in the uterus 100% of the activity was tPA. Only a small percentage of the OF proteolytic activity was plasminogen-dependent, probably due to the presence of PA inhibitors in this medium.
Jiménez Díaz
M
M
Instituto de Biología, Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Argentina.
Giunta
S
S
Valz-Gianinet
J
J
Pereyra-Alfonso
S
S
Flores
V
V
Miceli
D
D
D000581:Amidohydrolases / Q000378:metabolism; D000818:Animals; D001586:Benzoylarginine Nitroanilide / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002122:Calcium Chloride / Q000494:pharmacology; D002863:Chromogenic Compounds / Q000494:pharmacology; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D004848:Epithelium / Q000187:drug effects / Q000201:enzymology; D004950:Esterases / Q000378:metabolism; D005187:Fallopian Tubes / Q000187:drug effects / Q000201:enzymology*; D005260:Female; D006863:Hydrogen-Ion Concentration; D008647:Mesocricetus / Q000378:metabolism*; D010960:Plasminogen Activators / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D014106:Tosylarginine Methyl Ester / Q000378:metabolism; D014599:Uterus / Q000187:drug effects / Q000378:metabolism
D002863:Chromogenic Compounds; D015842:Serine Proteinase Inhibitors; D014106:Tosylarginine Methyl Ester; D001586:Benzoylarginine Nitroanilide; D004950:Esterases; D010960:Plasminogen Activators; D000581:Amidohydrolases; C019665:amidase; D002122:Calcium Chloride; D002118:Calcium