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A chemical genetics approach to examine the functions of AAA proteins.
- 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.
- Subjects :
- AAA Proteins antagonists & inhibitors
AAA Proteins ultrastructure
ATPases Associated with Diverse Cellular Activities genetics
ATPases Associated with Diverse Cellular Activities ultrastructure
Basic Helix-Loop-Helix Transcription Factors genetics
Endosomal Sorting Complexes Required for Transport genetics
Endosomal Sorting Complexes Required for Transport ultrastructure
Humans
Katanin ultrastructure
Microtubule-Associated Proteins ultrastructure
Microtubules genetics
Microtubules ultrastructure
Protein Conformation drug effects
Protein Domains genetics
Pyridines pharmacology
Triazoles chemistry
AAA Proteins genetics
Katanin genetics
Microtubule-Associated Proteins genetics
Pyridines chemistry
Subjects
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