在体内对孕激素受体DNA结合域的选择
Tholasi Nadhan Navien1, Ramesh Thevendran1, Marimuthu Citartan1
1Advanced Medical & Dental Institute (AMDI), Universiti Sains Malaysia, Bertam, 13200, Kepala Batas, Penang, Malaysia.
Analytical biochemistry
|December 24, 2024
概括
研究人员开发了一种新的DNA吸收体,PRDBDapt17,用于乳腺癌诊断中的孕激素受体检测. 这种体提供了抗体的替代品,显示出高的结合亲和力和广泛的类固醇激素受体应用的潜力.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 癌症的诊断 癌症的诊断
背景情况:
- 孕激素受体是乳腺癌诊断中的关键标记物,通常使用基于抗体的免疫组织化学检测.
- 抗体的局限性迫使人们探索其他分子识别工具,如体.
- 传统的SELEX对胺体的选择是严格的,这引起了人们对in silico方法的兴趣.
研究的目的:
- 执行针对孕激素受体DNA结合域 (PR DBD) 的DNA吸收体的in silico选择和实验验证.
- 为了利用从人类孕激素反应元素 (PREs) 获得的DNA序列来开发体.
- 评估阿普坦体作为乳腺癌和其他类固醇激素受体相关疾病的诊断工具的潜力.
主要方法:
- 用64个ssDNA序列的库对PR DBD进行DNA吸附体的in silico选择.
- 二级和三级结构确定,然后使用PatchDock进行对接模拟.
- 分子动态模拟和实验验证,使用直接的酶相关的阿普塔默测试 (ELASA).
主要成果:
- PRDBDapt17 emerged as the top candidate aptamer with the highest docking score (11334),由分子动力学证实,它是接得分 (11334) 的最佳候选亚胺酶之一. 通过分子动力学证实,PRDBDapt17成为了接得分 (11334) 的最佳候选亚胺酶之一.
- 直接ELASA验证显示,PRDBDapt17的检测极限为3.91nM.
- 平衡解离常数 (Kd) 确定为366.6nM,表明具有显著的结合亲和力.
- PRDBDapt17表现出与雌激素和雄激素受体的交叉反应性,这表明它有可能作为一种通用的类固醇激素受体结合剂.
结论:
- 在的方法成功地识别了PRDBDapt17,一个强大的DNA吸收器用于PR DBD识别.
- PRDBDapt17具有有利的检测极限和结合亲和力,适合用于诊断应用.
- 它作为类固醇激素受体的通用结合剂的潜力为在包括乳腺癌在内的激素相关疾病中更广泛的诊断实用性开辟了道路.
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