Isolation, purification, and characterization of mouse placental lactogen.
Isolation, purification, and characterization of mouse placental lactogen.
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小鼠胎盘催乳素的分离、纯化和表征。
DOI：
10.1073/pnas.79.3.771
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发表时间：
1982
影响因子：
11.1
通讯作者：
Talamantes,F
中科院分区：
文献类型：
--
作者：
Colosi,P;Marr,G;Lopez,J;Haro,L;Ogren,L;Talamantes,F
从妊娠14-18天的BALB/c小鼠胎盘中纯化了1840倍的胎盘催乳素，总收率为29%。纯化过程包括碱均质和提取，硫酸铵沉淀，Phenyl-Sepharose上的疏水相互作用色谱，CM-和DEAE-纤维素上的离子交换色谱，和Sephadex G-100上的凝胶排阻色谱。在10%碱性聚丙烯酰胺凝胶上，小鼠胎盘催乳素的Rf为0.19。在含有NaDodSO 4的凝胶中的电泳显示出具有对应于23，000 +/-1000的Mr的迁移率的单一条带。在8 M尿素/5%2-巯基乙醇中通过等电聚焦测定的等电点为7.1。当在鸽子作物囊试验中进行测试时，10微克小鼠胎盘催乳素产生的刺激与10微克绵羊催乳素引起的刺激相当。在兔乳腺放射受体测定中，小鼠胎盘催乳素比绵羊催乳素在从兔乳腺膜中置换125 I标记的绵羊催乳素方面的效力高150%。碘化纯化的小鼠胎盘催乳素可被小鼠胎盘催乳素、小鼠催乳素和绵羊催乳素从兔乳腺膜上置换。绵羊催乳素是小鼠胎盘催乳素的45%，它取代了兔乳腺膜上125 I标记的小鼠胎盘催乳素。小鼠胎盘催乳素与小鼠催乳素或小鼠生长激素的抗血清在放射免疫测定中无交叉反应。
Mouse placental lactogen was purified 1840-fold from BALB/c placentae from days 14-18 of gestation with an overall yield of 29%. The purification procedure included alkaline homogenization and extraction, ammonium sulfate precipitation, hydrophobic interaction chromatography on Phenyl-Sepharose, ion-exchange chromatography on CM- and DEAE-cellulose, and gel exclusion chromatography on Sephadex G-100. On 10% alkaline polyacrylamide gels, mouse placental lactogen had an Rf of 0.19. Electrophoresis in gels containing NaDodSO4 showed a single band with a mobility corresponding to a Mr of 23,000 +/- 1000. The isoelectric point, determined by isoelectric focusing in 8 M urea/5% 2-mercaptoethanol, was 7.1. When tested in the pigeon crop sac assay, 10 micrograms of mouse placental lactogen produced stimulation comparable with that evoked by 10 micrograms of ovine prolactin. In the rabbit mammary gland radioreceptor assay, mouse placental lactogen was 150% more potent than ovine prolactin in displacing 125I-labeled ovine prolactin from rabbit mammary gland membranes. Iodinated purified mouse placental lactogen could be displaced from rabbit mammary gland membranes by mouse placental lactogen, mouse prolactin, and ovine prolactin. Ovine prolactin was 45% as avid as mouse placental lactogen is displacing 125I-labeled mouse placental lactogen from rabbit mammary gland membranes. Mouse placental lactogen did not crossreact with antisera to mouse prolactin or mouse growth hormone in a radioimmunoassay.