Truncation of the porcine calcitonin receptor cytoplasmic tail inhibits internalization and signal transduction but increases receptor affinity.

Truncation of the porcine calcitonin receptor cytoplasmic tail inhibits internalization and signal transduction but increases receptor affinity.
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猪降钙素受体胞质尾部的截短会抑制内化和信号转导,但会增加受体亲和力。

DOI:
10.1210/mend.8.12.7708057
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发表时间:
1994
影响因子:
--
通讯作者:
P. Sexton
P. Sexton
中科院分区:
医学2区
文献类型:
--
作者:
D. Findlay;S. Houssami;H. Y. Lin;D. E. Myers;C. Brady;P. Darcy;K. Ikeda;T. J. Martin;P. Sexton

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一系列具有逐渐截短的羧基末端的突变型猪降钙素受体已在 COS 和 HEK 293 细胞中表达。所有形式的受体,包括完全缺乏细胞质尾部的受体,都能够结合 125I 标记的鲑鱼降钙素。然而,C 端结构域的去除导致受体的多种功能变化。首先,与野生型受体相比,鲑鱼降钙素对截短受体的结合亲和力增加,无论是在完整转染细胞中还是在细胞膜中测定。其次,对于被截短 44 或 83 个氨基酸的突变体,配体-受体复合物的内化大大减弱,但对于被截短 63 个氨基酸的中间形式则没有。第三,截短影响信号转导,对于猪降钙素受体来说,信号转导是通过细胞内cAMP和Ca2+的生成而发生的。对于内化缺陷的相同突变体,腺苷酸环化酶反应的强度大大降低。在每种受体形式的表达大致相等的条件下,C 末端截短会降低细胞内 Ca2+ 反应的强度。这些结果引起人们对猪降钙素受体细胞质尾部功能重要性的关注,并表明羧基末端与其他受体结构域和/或细胞调节元件之间的分子内相互作用。
A series of mutant porcine calcitonin receptors with progressively truncated carboxy termini have been expressed in COS and HEK 293 cells. All forms of the receptor, including those totally lacking the cytoplasmic tail, were able to bind 125I-labeled salmon calcitonin. However, removal of C-terminal domains resulted in multiple functional changes in the receptor. First, compared with the wild type receptor, affinity of binding of salmon calcitonin was increased for truncated receptors, whether determined in intact transfected cells or in cell membranes. Second, internalization of the ligand-receptor complex was greatly attenuated for mutants truncated by 44 or 83 amino acids but not for an intermediate form truncated by 63 amino acids. Third, truncation affected signal transduction, which for the porcine calcitonin receptor occurs by generation of intracellular cAMP and Ca2+. The magnitude of adenylate cyclase responses was much reduced for the same mutants defective in internalization. Under conditions where expression of each receptor form was approximately equal, the magnitude of intracellular Ca2+ responses was decreased by C-terminal truncation. These results draw attention to the functional significance of the cytoplasmic tail of the porcine calcitonin receptor and suggest intramolecular interactions between the carboxy terminus and other receptor domains and/or cellular regulatory elements.
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影响因子: 4
作者:
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