Motor mechanism of native ATPase complexes

AAA+超家族的ATPase(与各种细胞活动相关的ATPase)通常是多蛋白复合体的核心，负责以能量依赖的方式及时破坏或重塑大分子。它们构成了最大的蛋白质超家族之一，是细胞内蛋白质、膜或DNA大多数重组和循环过程中的关键组成部分。它们独特的功能特异性来自于将高度保守的AAA+结构域与辅助结构域和蛋白质结合在一起，这有助于AAA+活性的微调。本项目的目标是确定不同功能的AAA+组装的三维结构，以了解AAA+蛋白共同的潜在作用机制及其辅助因子的调节。使用单粒子低温电子显微镜将探索(I)具有不同大小结构域插入的热休克蛋白(HSP)100变异体、(Ii)蛋白酶体亚复合体和(Iii)异寡聚过氧化体ATPase组件的功能状态。

ATPases of the AAA+ superfamily (ATPases associated with various cellular activities) are often at the core of multi-protein complexes responsible for timely destruction or remodeling of macromolecules in an energy dependent manner. They form one of the largest protein superfamilies and are crucial components in most re-organization and recycling processes of protein, membrane or DNA within the cell. Their unique functional specificity derives from combining the highly conserved AAA+ domain with accessory domains and proteins which facilitate fine tuning of the AAA+ activity. The goal of this project is to determine the three dimensional structure of different functional AAA+ assemblies in order to understand the common underlying mechanism of action of AAA+ proteins and its regulation by accessory factors. Using single particle cryo electron microscopy the functional states of (i) heat shock protein (Hsp) 100 variants with varying sized domain insertions, (ii) proteasomal subcomplexes and (iii) hetero oligomeric peroxisomal ATPase assemblies will be explored.