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A chemical genetics approach to examine the functions of AAA proteins.

Authors :
Cupido T
Jones NH
Grasso MJ
Pisa R
Kapoor TM
Source :
Nature structural & molecular biology [Nat Struct Mol Biol] 2021 Apr; Vol. 28 (4), pp. 388-397. Date of Electronic Publication: 2021 Mar 29.
Publication Year :
2021

Abstract

The structural conservation across the AAA (ATPases associated with diverse cellular activities) protein family makes designing selective chemical inhibitors challenging. Here, we identify a triazolopyridine-based fragment that binds the AAA domain of human katanin, a microtubule-severing protein. We have developed a model for compound binding and designed ASPIR-1 (allele-specific, proximity-induced reactivity-based inhibitor-1), a cell-permeable compound that selectively inhibits katanin with an engineered cysteine mutation. Only in cells expressing mutant katanin does ASPIR-1 treatment increase the accumulation of CAMSAP2 at microtubule minus ends, confirming specific on-target cellular activity. Importantly, ASPIR-1 also selectively inhibits engineered cysteine mutants of human VPS4B and FIGL1-AAA proteins, involved in organelle dynamics and genome stability, respectively. Structural studies confirm our model for compound binding at the AAA ATPase site and the proximity-induced reactivity-based inhibition. Together, our findings suggest a chemical genetics approach to decipher AAA protein functions across essential cellular processes and to test hypotheses for developing therapeutics.

Details

Language :
English
ISSN :
1545-9985
Volume :
28
Issue :
4
Database :
MEDLINE
Journal :
Nature structural & molecular biology
Publication Type :
Academic Journal
Accession number :
33782614
Full Text :
https://doi.org/10.1038/s41594-021-00575-9