Afting E G, Becker M L, Elce J S
Biochem J. 1979 Jan 1;177(1):99-106. doi: 10.1042/bj1770099.
A supernatant fraction was prepared from rat uterine myometrium by homogenization, sonication and centrifugation. In this supernatant the protein concentration and the activities of an acid proteinase, an acid phosphatase and a proteinase inhibitor were measured. From the fibrous sediment, after washing with 0.5% Triton X-100 and with water, an actomyosin-containing solution was obtained by extraction with 0.6M-NaCl, and in this extract the protein concentration and a neutral proteinase activity were measured. The myometrial wet weight and the activities of the acid proteinase, acid phosphatase and proteinase inhibitor increased by factors of 3-15 during pregnancy and decreased to the same or a greater extent during involution. The amount of protein extracted with 0.6M-NaCl increased by a factor of only 2.3 and the neutral proteinase activity remained essentially constant during pregnancy and involution. The pH optimum of the neutral proteinase, and its pattern of activity compared with those of the lysosomal enzymes, show that the neutral proteinase is not of lysosomal origin. Actomyosin is degraded by the neutral proteinase activity in vitro. Since actomyosin is rapidly broken down only after parturition, the action of the neutral proteinase activity on actomyosin, if this occurs in vivo, must be regulated in some way. The proteinase-inhibitor activity measured in the first supernatant varied in a manner which suggested that it could be involved in this control.
通过匀浆、超声处理和离心从大鼠子宫肌层制备上清液部分。在该上清液中测量蛋白质浓度以及酸性蛋白酶、酸性磷酸酶和蛋白酶抑制剂的活性。从纤维状沉淀物中,先用0.5% Triton X - 100洗涤,再用水洗涤,然后用0.6M - NaCl提取得到含肌动球蛋白的溶液,在该提取物中测量蛋白质浓度和中性蛋白酶活性。在怀孕期间,子宫肌层湿重以及酸性蛋白酶、酸性磷酸酶和蛋白酶抑制剂的活性增加3 - 15倍,在 involution期间下降到相同或更大程度。用0.6M - NaCl提取的蛋白质量仅增加2.3倍,中性蛋白酶活性在怀孕和 involution期间基本保持恒定。中性蛋白酶的最适pH值及其与溶酶体酶相比的活性模式表明,中性蛋白酶并非源自溶酶体。肌动球蛋白在体外被中性蛋白酶活性降解。由于肌动球蛋白仅在分娩后才迅速分解,因此如果中性蛋白酶活性在体内作用于肌动球蛋白,其作用必定以某种方式受到调节。在第一个上清液中测量的蛋白酶抑制剂活性变化方式表明它可能参与了这种调节。