Endogenous blockade of 1,25-dihydroxyvitamin D-receptor binding in New World primate cells.
Endogenous blockade of 1,25-dihydroxyvitamin D-receptor binding in New World primate cells.
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新大陆灵长类细胞中 1,25-二羟基维生素 D 受体结合的内源性阻断。
DOI：
10.1172/jci115108
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Adams,JS
中科院分区：
文献类型：
--
作者：
Gacad,MA;Adams,JS
当通过1，25-二羟维生素D3（1，25（OH）2-D3）-受体（VDR）结合分析或1，25（OH）2-D3-VDR导向的生物反应性进行评估时，与旧世界灵长类（卡他鼻）细胞相比，来自一些新世界灵长类（阔鼻）的培养细胞显示VDR的可变降低。为了研究灵长类动物中VDR表达的差异，我们对培养的皮肤成纤维细胞中的VDR进行了免疫印迹分析，这些成纤维细胞来自Pithirrhine属和Aotus属的阔鼻类动物和Presbytis属的卡他类动物;虽然是阔鼻类动物，但猫头鹰猴（Aotus）表达了卡他类动物（野生型）表型的VDR。尽管通过配体结合分析，来自三个原型灵长类属的细胞之间的VDR含量存在10倍差异，但通过免疫印迹分析检测到的50 kD VDR稳态水平差异小于或等于10%细胞提取物。我们研究了这种明显的差异，在免疫印迹和配体结合分析的VDR含量混合VDR含有核提取物的等效蛋白质浓度从各种灵长类动物。将Pithelium和Aotus成纤维细胞提取物与Presbytis提取物共孵育，分别使混合物中的特异性1，25（OH）2-D3结合减少了90%和95%。通过混合猫头鹰猴细胞系OMK和维生素D抗性绒猴B淋巴母细胞系B 95 -8的核提取物，获得了类似的结果。B 95 -8提取物经胰蛋白酶或热处理后，混合物中野生型1，25（OH）2-D3结合谱得以恢复。这些数据表明，一些新世界灵长类动物细胞含有可溶性蛋白，可阻止细胞内1，25（OH）2-D3-VDR结合。这种蛋白质表达的数量差异可能是新世界灵长类动物中1，25（OH）2-D3和其他严重程度不同的类固醇激素抵抗状态的原因。
When assessed by 1,25-dihydroxyvitamin D3 (1,25(OH)2-D3)-receptor (VDR) binding analysis or 1,25(OH)2-D3-VDR-directed bioresponsiveness, cultured cells from some New World primates (platyrrhines) demonstrate a variable decrement in VDR when compared with Old World primate (catarrhine) cells. To study this difference in VDR expression among primates, we performed immunoblot analysis of the VDR in cultured dermal fibroblasts from platyrrhines in the genera Pithecia and Aotus and from catarrhines in the genus Presbytis; although a platyrrhine, the owl monkey (Aotus) expresses a VDR of the catarrhine (wild type) phenotype. Despite a 10-fold difference in the content of VDR by ligand binding analysis among cells from the three prototypic primate genera, there was a less than or equal to 10% difference in the steady-state level of 50-kD VDR detected by immunoblot analysis of cellular extracts. We investigated this apparent discrepancy in the content of VDR in immunoblots and ligand binding analyses by mixing VDR-containing nuclear extracts of equivalent protein concentration from the various primates. Coincubation of Pithecia and Aotus fibroblast extracts with Presbytis extract diminished specific 1,25(OH)2-D3 binding in the mix by 90% and 95% respectively. Similar results were obtained by mixing nuclear extracts of the owl monkey cell line, OMK, and the vitamin D resistant marmoset B-lymphoblast cell line B95-8. A wild type 1,25(OH)2-D3-binding profile was restored in mixtures after trypsin or heat treatment of the B95-8 extract. These data indicate that some New World primate cells contain a soluble protein that prevents intracellular 1,25(OH)2-D3-VDR binding. It is possible that the quantitative differences in the expression of this protein are responsible for 1,25(OH)2-D3 and other steroid hormone resistant states of variable severity in New World primates.Images