Jiménez Díaz M, Giunta S, Valz-Gianinet J, Pereyra-Alfonso S, Flores V, Miceli D
Instituto de Biología, Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Argentina.
Mol Reprod Dev. 2000 Jan;55(1):47-54. doi: 10.1002/(SICI)1098-2795(200001)55:1<47::AID-MRD7>3.0.CO;2-J.
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.
目前，大多数输卵管蛋白的生理作用仍不清楚。在这项研究中，我们提供证据表明，输卵管分泌物以及粗制的输卵管组织提取物显示出类蛋白水解酯酶和酰胺酶活性。超排仓鼠雌性输卵管中输卵管酶的蛋白水解活性高于正常仓鼠，这表明促性腺激素会刺激这些酶的合成和分泌。在输卵管组织提取物（OE）和输卵管液（OF）的15,600-g上清液中分析了它们的一些特性。对合成底物对甲苯磺酰-L-精氨酸甲酯盐酸盐（TAME）和α-N-苯甲酰-DL-精氨酸对硝基苯胺盐酸盐（BAPNA）的酶活性被钙离子激活，在pH 7.5时达到最大值，并被大豆胰蛋白酶抑制剂（SBTI）、N-α-对甲苯磺酰-L-赖氨酸氯甲基酮盐酸盐（TLCK）、苯甲基磺酰氟（PMSF）和苯甲脒抑制。经WGA-琼脂糖亲和柱识别的OE糖蛋白组分（占总蛋白的37%）显示出与OE和OF酶性质相似的蛋白水解活性。蛋白酶活性可归因于在经Triton X-100处理的组织粗膜组分（Triton-CMF）和超排雌性的输卵管分泌物中检测到的纤溶酶原激活剂（PA）。在Triton-CMF组分中，100%的蛋白水解活性依赖于纤溶酶原。使用氨氯吡咪（一种选择性尿激酶型纤溶酶原激活剂（uPA）抑制剂）表明，这种活性的90%归因于组织型纤溶酶原激活剂（tPA），10%归因于uPA，而在子宫中100%的活性是tPA。OF的蛋白水解活性只有一小部分依赖于纤溶酶原，这可能是由于该介质中存在PA抑制剂。
