AI Insight
Researchers developed new chemical tools to study and control ATAT1, an enzyme that marks stable microtubules by adding acetyl groups to tubulin proteins. They engineered a mutant version of ATAT1 that can attach clickable chemical tags to tubulin, enabling visualization of modified microtubules through fluorescence imaging. Additionally, they created an inhibitor molecule that can suppress both natural and engineered modifications by ATAT1.
Why it matters
These tools enable scientists to directly observe and manipulate microtubule modifications in living cells, which could advance understanding of cellular processes like cell division, intracellular transport, and diseases involving microtubule dysfunction. The clickable labeling method provides a precise way to track specific protein modifications that was previously unavailable.
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⚠️ Preprint – Noch nicht peer-reviewed
Dieser Artikel wurde noch nicht von unabhängigen Experten begutachtet. Die Ergebnisse sind vorläufig und sollten mit Vorsicht interpretiert werden.
Acetylation of -tubulin K40 by -tubulin acetyltransferase 1 (ATAT1) using acetyl-coenzyme A (Ac-CoA) marks stable microtubule populations, yet chemical tools to directly measure ATAT1 ligand engagement, inhibit its activity, or visualize ATAT1-mediated modification on intact microtubules remain limited. Through the development of a quantitative binding assay, we uncovered that ATAT1 can bind unnatural cofactors but fails to efficiently use them in acyl-transfer reactions. Structure-guided mutation subsequently yielded ATAT1-L163A, which successfully installed clickable handles at the native -tubulin K40 site of synthetic tubulin peptides, -tubulin, and intact microtubules. Cu(I)-catalyzed azide-alkyne cycloaddition enabled visualization of modified microtubules by in-gel fluorescence and microscopy. Moreover, we report a p11-CoA bisubstrate inhibitor that suppressed both native acetylation and engineered acylation. Together, these tools provide chemically controlled access to ATAT1 activity and a site-verified, clickable K40 modification on intact microtubules.
Source: Chemical Interrogation and Reprogramming of ATAT1-Mediated Tubulin Acetylation