Structural and mechanistic insights into Hsp104 function revealed by synchrotron X-ray footprinting.

ABSTRACT: Hsp104 is a hexameric AAA⁺ ring translocase, which drives protein disaggregation in nonmetazoan eukaryotes. Cryo-EM structures of Hsp104 have suggested potential mechanisms of substrate translocation, but precisely how Hsp104 hexamers disaggregate proteins remains incompletely understood. Here, we employed synchrotron X-ray footprinting to probe the solution-state structures of Hsp104 monomers in the absence of nucleotide and Hsp104 hexamers in the presence of ADP or ATPγS (adenosine 5'-O-(thiotriphosphate)). Comparing side-chain solvent accessibilities between these three states illuminated aspects of Hsp104 structure and guided design of Hsp104 variants to probe the disaggregase mechanism in vitro and in vivo We established that Hsp104 hexamers switch from

SUBMITTER: Sweeny EA 

PROVIDER: S-EPMC7008382 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature