与Dictyostelium--a dipeptide模型系统的Skp1中的Pro(143) 的翻译后修改相关的形状变化
Chamini V Karunaratne1, Thomas K Weldeghiorghis, Christopher M West
1Department of Chemistry, Louisiana State University , Baton Rouge, Louisiana 70803, United States.
Journal of the American Chemical Society
|September 25, 2014
概括
翻译后的修改,如prolyl氧化和糖化,改变了蛋白质的结构. 这些Skp1的变化,SCF泛基因酶的子单元,影响氧气感应和蛋白质相互作用.
科学领域:
- 生物化学 生物化学
- 结构生物学 结构生物学
- 分子生物学分子生物学
背景情况:
- 在SCF中,乌比基酸酶子单元Skp1经历了prolyl氧化和糖化.
- 这些修改影响了Skp1构造,F盒蛋白相互作用和生物体的氧气传感.
- 了解这些变化的分子基础至关重要.
研究的目的:
- 调查通过什么分子机制的prolyl氧化和糖基化影响Skp1形状和键动态.
- 阐明翻译后修改在调节蛋白质结构和功能的作用.
主要方法:
- 合成各种修饰的终端盖的Thr-Pro二.
- 核磁共振 (NMR) 谱学,包括 (1) H 和 (13) C 核磁共振,用于分析形状和化学变化.
- 磁化转移实验以确定cis-trans异构化反应速率.
- 核重整效应 (NOE) 实验用于评估空间接近.
主要成果:
- 观察到对转变变形的偏好越来越多,顺序如下:Pro < 基 (Hyp) < 糖化基 ([α-(1,4) GlcNAc]Hyp).
- 林的氧化降低了转化成cis的转化率,并增加了转化成cis的转化率.
- NOE实验表明,N端Thr残留的侧链影响了糖部分远离罗利丁环的方向.
结论:
- 在Skp1中对林残留物的翻译后修改显著改变了键构造和异构化动态.
- 这些构造变化为Skp1功能和全长蛋白质中氧气传感的影响提供了一个分子机制.
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