Jaffe R C, Donnelly K M, Mavrogianis P A, Verhage H G
Department of Physiology, University of Illinois, Chicago 60680.
Mol Endocrinol. 1989 Nov;3(11):1807-14. doi: 10.1210/mend-3-11-1807.
In the cat, a group of low molecular weight secretory proteins have previously been shown to appear in the endometrium after progesterone (P) administration to an estrogen (E2)-primed animal. Using a polyclonal antibody to these progesterone-dependent proteins (PDP) we have isolated a recombinant cDNA clone corresponding to the mRNA for PDP from a cDNA library prepared using poly(A+) RNA from the endometrium of P-treated E2-primed cats. Comparison of Western blots using the polyclonal antibody and epitope selected antibody demonstrated that the multiple molecular weight and isoelectric forms of the PDP are immunologically related and potentially products of the same gene. Northern analysis revealed that the mRNA for the PDP in the endometrium of P-treated E2-primed cats was 1.8 kilobases in length. Using slot blot analysis, we found that the PDP mRNA levels were low in the endometrium of ovariectomized animals and undetectable in E2-treated animals. With 1 day of P treatment the PDP mRNA levels were readily detectable and they peaked after 5 to 7 days of P treatment. No PDP mRNA was detectable in myometrium, oviduct, or ovary. Sequence analysis revealed that PDP had significant homology to human, rat, and mouse cathepsin L at the nucleotide (80%, 74%, and 73%, respectively) and amino acid (68%, 65%, and 63%, respectively) level. We suggest that PDP via its collagenolytic and elastolytic activities as a cathepsin L is responsible for preparing the endometrium for blastocyst implantation.
在猫中，先前已表明，给用雌激素（E2）预处理的动物注射孕酮（P）后，一组低分子量分泌蛋白会出现在子宫内膜中。利用针对这些孕酮依赖性蛋白（PDP）的多克隆抗体，我们从一个使用经P处理的E2预处理猫子宫内膜的聚腺苷酸加尾（poly(A+)）RNA制备的cDNA文库中，分离出了一个与PDP的mRNA相对应的重组cDNA克隆。使用多克隆抗体和表位选择抗体进行的蛋白质免疫印迹比较表明，PDP的多种分子量和等电形式在免疫上相关，并且可能是同一基因的产物。Northern印迹分析显示，经P处理的E2预处理猫子宫内膜中PDP的mRNA长度为1.8千碱基。通过狭缝印迹分析，我们发现去卵巢动物的子宫内膜中PDP mRNA水平较低，而在E2处理的动物中则检测不到。给予1天的P处理后，PDP mRNA水平易于检测到，并且在P处理5至7天后达到峰值。在子宫肌层、输卵管或卵巢中未检测到PDP mRNA。序列分析表明，PDP在核苷酸水平（分别为80%、74%和73%）和氨基酸水平（分别为68%、65%和63%）与人类、大鼠和小鼠组织蛋白酶L具有显著同源性。我们认为，作为组织蛋白酶L的PDP通过其胶原分解和弹性蛋白分解活性，负责为胚泡着床准备子宫内膜。
