Purification and partial characterization of relaxin and relaxin precursors from the hamster placenta.
Purification and partial characterization of relaxin and relaxin precursors from the hamster placenta.
复制标题
仓鼠胎盘松弛素和松弛素前体的纯化和部分表征。
DOI：
10.1095/biolreprod49.1.154
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发表时间：
1993
影响因子：
3.6
通讯作者：
Chalovich,JM
中科院分区：
文献类型：
--
作者：
Renegar,RH;Owens,CR;Chalovich,JM
以往的免疫学研究表明，仓鼠松弛蛋白的分子结构与猪松弛蛋白有很大的不同。本研究从金黄地鼠胎盘中分离纯化了松弛蛋白，并对其生化性质进行了研究。取孕14天和15天的胎盘在0.26N盐酸-62.5%丙酮中匀浆。离心后，丙酮沉淀出可溶性蛋白质。可溶性蛋白质用羧甲基纤维素离子交换柱，结合蛋白用0.1和0.3M的氯化钠洗脱。Western印迹分析在0.1M氯化钠洗脱液中检测到16.5、18.7和36.0 kDa松弛蛋白免疫反应(IR)蛋白，在0.3M氯化钠洗脱液中检测到5.6 kDa松弛蛋白免疫反应(IR)蛋白。经Sephadex G-50凝胶过滤、离子交换高效液相色谱(IEC)和C18-高效液相色谱(C18-HPLC)纯化得到5.6 kDa蛋白。在电泳前还原5.6-kDa蛋白导致了一条低分子量的单一条带，这表明仓鼠松弛蛋白由两条分子质量大致相等的链组成。5.6 kDa蛋白的等电点为7.78。通过凝胶过滤和离子交换高效液相色谱分离纯化得到16.5 kDa和18.7 kDa的IR蛋白。观察到16.5 kDa和18.7 kDa蛋白质至少有5个等电点变异。松弛蛋白-IR蛋白5.6和18.7的N端氨基酸为精氨酸，随后的循环显示一个与其他物种的松弛蛋白完全一致的部分序列。
Previous immunological studies have indicated that the molecular structure of hamster relaxin is quite different from that of porcine relaxin. In the present study, hamster relaxin was purified from placentas and characterized in order to investigate its biochemical properties. Placentas from Days 14 and 15 of gestation were homogenized in 0.26 N HCl-62.5% acetone containing protease inhibitors. After centrifugation, soluble proteins were acetone precipitated. Soluble proteins were applied to a carboxymethyl cellulose ion-exchange column and bound proteins were eluted with 0.1 and 0.3 M NaCl. Western blot analysis detected 16.5-, 18.7-, and 36.0-kDa relaxin-immunoreactive (IR) proteins within the 0.1 M NaCl eluant and detected a 5.6-kDa relaxin-IR protein within the 0.3 M NaCl eluant. The 5.6-kDa protein was purified to homogeneity by gel filtration (Sephadex G-50), ion-exchange HPLC, and C18 -HPLC. Reduction of the 5.6-kDa protein prior to electrophoresis resulted in a single band of lower molecular mass, suggesting that hamster relaxin consists of two chains of approximately equal molecular mass. Isoelectric point of the 5.6-kDa protein was 7.78. The 16.5- and 18.7-kDa IR proteins were copurified by gel filtration and ion-exchange HPLC. At least five isoelectric point variants were observed for the 16.5- and 18.7-kDa proteins. The N-terminal amino acid for the 5.6 and 18.7 relaxin-IR proteins was arginine, and subsequent cycles indicated an identical partial sequence that was consistent with that for relaxins from other species.
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怀孕母猪卵巢中高分子量形式的松弛素。
DOI：
--
发表时间：
1978
期刊：
Biochemical and Biophysical Research Communications - BBRC
影响因子：
--
作者：
S. Kwok;W. Chamley;G. Bryant
通讯作者：
G. Bryant
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影响因子：
4.8
作者：
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通讯作者：
O. Sherwood
影响因子：
4.8
作者：
D. Stewart;B. Nevins;É. Hadas;R. Vandlen
通讯作者：
D. Stewart;B. Nevins;É. Hadas;R. Vandlen
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影响因子：
4.8
作者：
B. Steinetz;E. O'Byrne;L. Goldsmith;M. B. Anderson
通讯作者：
M. B. Anderson
金仓鼠胎盘巨型滋养层细胞的超微结构形态和松弛素免疫定位。
影响因子：
--
作者：
T. Johns;R. Renegar
通讯作者：
R. Renegar