Covalent modification of proteins by ligands of steroid hormone receptors.
Covalent modification of proteins by ligands of steroid hormone receptors.
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类固醇激素受体配体对蛋白质的共价修饰。
DOI：
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发表时间：
1992
影响因子：
11.1
通讯作者：
T. Breitman
中科院分区：
文献类型：
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作者：
N. Takahashi;T. Breitman
视黄酰化，视黄酸（RA）酰化，是发生在各种真核细胞系中的蛋白质的共价修饰。在本研究中，我们发现HL-60细胞中的蛋白质被17 β-[3 H]雌二醇（E2）、[3 H]孕酮（Pg）、1 α，25-二羟基[3 H]维生素D3 [1，25（OH）2D 3]、[125 I]三碘甲状腺原氨酸（T3）、[125 I]甲状腺素（T4）和[3 H]前列腺素E2（PGE 2）标记。除PGE 2外，所有这些激素都是类固醇激素受体家族的配体。此外，5 μ M酮康唑，细胞色素P450依赖性酶的抑制剂的生长培养基中，增加了约2倍的T3，T4，1，25（OH）2D 3和PGE 2的蛋白质标记。与此相反，酮康唑并没有显着改变的程度标记RA，E2，或PG。碱性甲醇分解，裂解酯键，释放可变百分比的放射性配体结合蛋白质。这些值对于RA和PGE 2约为80%;对于T3、T4和Pg约为50%;对于E2和1，25（OH）2D 3约为20%。用硫醚裂解试剂、碘甲烷或雷尼镍催化剂处理，释放< 2%的共价结合的配体。每个配体的标记蛋白质的二维聚丙烯酰胺凝胶电泳模式是独特的。M（r）47，000和51，000的蛋白质被RA、E2、T3和T4标记。这些蛋白质具有与RI和RII相同的迁移率，RI和RII是I型和II型cAMP依赖性蛋白激酶的cAMP结合调节亚基。1，25（OH）2D 3也与M（r）47，000和51，000的蛋白质结合。然而，这些蛋白质具有不同于RI或RII的pI值。这些结果表明，类固醇激素受体家族和PGE 2的配体的一些活动可能是通过它们的蛋白质的共价修饰介导的。
Retinoylation, acylation with retinoic acid (RA), is a covalent modification of proteins occurring in a variety of eukaryotic cell lines. In this study, we found that proteins in HL-60 cells were labeled by 17 beta-[3H]estradiol (E2), [3H]progesterone (Pg), 1 alpha,25-dihydroxy[3H]vitamin D3 [1,25(OH)2D3], [125I]triiodothyronine (T3), [125I]thyroxine (T4), and [3H]prostaglandin E2 (PGE2). All of these hormones, except PGE2, are ligands of the steroid hormone receptor family. Addition to the growth medium of 5 microM ketoconazole, an inhibitor of cytochrome P450-dependent enzymes, increased about 2-fold the labeling of proteins by T3, T4, 1,25(OH)2D3, and PGE2. In contrast, ketoconazole did not change markedly the extent of labeling by RA, E2, or Pg. Alkaline methanolysis, which cleaves ester bonds, released variable percentages of the radioactive ligands bound to protein. These values were about 80% for RA and PGE2; 50% for T3, T4, and Pg; and 20% for E2 and 1,25(OH)2D3. Treatment with thioether-cleavage reagents, iodomethane or Raney nickel catalyst, released < 2% of the covalently bound ligands. Two-dimensional polyacrylamide gel electrophoresis patterns of labeled proteins were unique for each ligand. Proteins of M(r) 47,000 and 51,000 were labeled by RA, E2, T3, and T4. These proteins had the same mobilities as RI and RII, the cAMP-binding regulatory subunits of type I and type II cAMP-dependent protein kinases. 1,25(OH)2D3 also bound to proteins of M(r) 47,000 and 51,000. However, these proteins had pI values different from those of RI or RII. These results suggest that some activities of ligands of the steroid hormone receptor family and of PGE2 may be mediated by their covalent modification of proteins.