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人类干扰素调节因子6的N端域的生物物理和功能特征

Binita Kumari Sinha¹, Devbrat Kumar², Priyabrata Meher¹

¹Department of Life Science, School of Earth, Biological and Environmental Sciences, Central University of South Bihar, Gaya, Bihar, 824236, India.

Molecular biology reports

|March 1, 2024

概括

干扰素调节因子6的N端域 (NTD-IRF6) 主要是α螺旋的,在pH值9.0时非常稳定. 这个域特别结合VRE-IFNβ促进子,具有高度亲和力,对于调节基因表达至关重要.

科学领域:

分子生物学分子生物学
蛋白质生物化学蛋白质生物化学
遗传学是一个遗传学.

背景情况:

干扰素调节因子6 (IRF6) 在胚胎发育过程中对 palatal 融合至关重要.
在IRF6的突变与 orofacial裂障碍有关.
IRF6的N端域 (NTD) 在调节基因表达方面发挥作用,包括IFNβ.

研究的目的:

描述IRF6 (NTD-IRF6) 的N端域的蛋白质折叠和稳定性.
研究NTD-IRF6对VRE-IFNβ促进子区域的DNA结合活性和亲和力.

主要方法:

用于域边界识别的in silico分析.
亚克隆,过度表达和净化复合NTD-IRF6.
循环二极化 (CD) 和内在光谱学用于蛋白质折叠研究.
凝阻滞测定和生物层干扰测量 (BLI) 用于DNA结合分析.

主要成果:

NTD-IRF6成功地被过度表达并净化为GST融合蛋白.
CD研究表明主要含有α-螺旋体,在pH值9.0时具有最大稳定性.
光光谱学显示,托残留物主要埋在蛋白质内.
凝延迟和BLI证明了对VRE-IFNβ促进体具有高亲和力 (K_D = 7.96 × 10-10 M) 的特定DNA结合.

结论:

NTD-IRF6 呈现出混合的二次结构 (α-螺旋和β-叶片),其展开取决于温度.
蛋白质对尿素诱导的展开表现出适度的耐受性.
NTD-IRF6与VRE-IFNβ促进区具有特定和高亲和度的相互作用.

关键词:

生物层干涉测量 (BIOLAYER INTERFEROMETRY) 是一种干涉测量方法.一个圆形的二重化.的DNA结合域域.在EMSA上,它是EMSA.内在的光是一种内在的光.净化净化净化净化

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