Solubilization of a guanine nucleotide-sensitive parathyroid hormone-receptor complex from canine renal cortex.
Solubilization of a guanine nucleotide-sensitive parathyroid hormone-receptor complex from canine renal cortex.
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犬肾皮质鸟嘌呤核苷酸敏感的甲状旁腺激素受体复合物的溶解。
DOI：
10.1007/978-1-4684-5206-8_41
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发表时间：
1986
影响因子：
--
通讯作者：
Arnaud,CD
中科院分区：
文献类型：
--
作者：
Nissenson,RA;Mann,E;Winer,J;Teitelbaum,A;Arnaud,CD
犬肾皮质PTH受体与激动剂配体占据膜相关受体后被溶解[125I]牛(b) PTH-(1-34)。为了在增溶过程中稳定结合，需要使用高浓度的牛血清白蛋白（最佳浓度为5%）和适当的洗涤剂（0.5% 3-[（3-胆酰胺丙基）二甲氨基]2-羟基-1-原-全磺酸盐，0.5% 3-[（3-胆酰胺丙基）二甲氨基]1-丙磺酸盐，或0.5-1.0%洋地黄苷）。可溶性部分（240,000 ×gavsupernatant）含有与大分子组分相关的[125I]bPTH-(1-34)以及在溶解过程中解离的未结合的[125I]bPTH-(1-34)。可溶性大分子复合物具有由[125I]bPTH-(1-34)受体刺激鸟嘌呤核苷酸结合蛋白（N8）组成的三元复合物的功能特性。因此，标记的甲状旁腺素在30℃时解离缓慢（t½= 75 min）；在GTP存在的情况下（10−4m）， 75%的位点表现出快速解离动力学（t½= 2.3 min）。该效应具有核苷酸特异性，即GTP≃GTPγ s≃GDPβ s > ITP≃guananylylimidodiphosphate→GMP≃App(NH)p。ATP无效。GTP在浓度为200 nm时产生半最大响应。这些结果与报道的纯化N8的核苷酸特异性和亲和力一致。在结合反应中，用n -乙基马来酰亚胺处理膜使溶解的配合物对GTP难溶。凝胶过滤层析（Sepharose 6B）发现了一个gtp敏感复合物，该复合物被洗脱在180000道尔顿的无洗涤剂球形蛋白的预期位置。该复合物可能由60,000至70,000道尔顿的pth结合亚基（先前通过光亲和标记确定）和N8组成。(endocrinology118:932 - 939, 1986)
Canine renal cortical PTH receptors were solubilized after occupancy of membrane-associated receptors with the agonist ligand [125I]bovine (b) PTH-(1–34). Stabilization of binding during solubilization required the use of high concentrations of BSA (optimally 5%) and appropriate detergents (0.5% 3-[(3-cholamidopropyl)dimethylammonio]2-hydroxy-1-pro-panesulfonate, 0.5% 3-[(3-cholamidopropanyl)dimethylam-monio]1-propanesulfonate, or 0.5–1.0% digitonin). The soluble fraction (240,000 ×gavsupernatant) contained [125I]bPTH-(1–34) associated with macromolecular components as well as unbound [125I]bPTH-(1–34) that dissociated during solubilization. The soluble macromolecular complex had functional properties expected of a ternary complex consisting of [125I]bPTH-(1–34) receptor stimulatory guanine nucleotide-binding protein (N8). Thus, the dissociation of labeled PTH at 30 C was slow (t½= 75 min); in the presence of GTP (10−4m), 75% of the sites displayed rapid dissociation kinetics (t½= 2.3 min). This effect was nucleotide specific, with GTP ≃ GTPγS ≃ GDP > GDPβS > ITP ≃ guanylylimidodiphosphate ≫ GMP ≃ App(NH)p. ATP was ineffective. GTP produced a half-maximal response at a concentration of 200 nm. These results are consistent with the reported nucleotide specificity and affinity of purified N8. Treatment of membranes withN-ethylmaleimide during the binding reaction rendered the solubilized complex refractory to GTP. Gel filtration chromatography (Sepharose 6B) revealed a GTP-sensitive complex that eluted in the position expected of a detergent-free spherical protein of 180,000 daltons. This complex may consist of the 60,000 to 70,000-dalton PTH-binding subunit (previously identified by photoaffinity labeling) together with N8. (Endocrinology118:932–939, 1986)
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影响因子：
64.8
作者：
R. Cerione;B. Strulovici;J. Benovic;R. Lefkowitz;M. Caron
通讯作者：
M. Caron
禽类骨细胞中的甲状旁腺激素受体。
DOI：
10.1073/pnas.79.6.2061
发表时间：
1982
影响因子：
11.1
作者：
Pliam,NB;Nyiredy,KO;Arnaud,CD
通讯作者：
Arnaud,CD
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DOI：
10.1097/00006254-198102000-00017
发表时间：
1980
期刊：
The New England journal of medicine
影响因子：
--
作者：
Z. Farfel;A. Brickman;H. Kaslow;H. Bourne
通讯作者：
H. Bourne
影响因子：
4.8
作者：
A. Teitelbaum;G. Strewler
通讯作者：
A. Teitelbaum;G. Strewler
哺乳动物肾脏质膜蛋白的表征。
影响因子：
4.8
作者：
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通讯作者：
E. Landon