6NYV

Structure of spastin AAA domain in complex with a quinazoline-based inhibitor

Summary for 6NYV

• Entry DOI: 10.2210/pdb6nyv/pdb
• Related: 6NYW
• Descriptor: Spastin, SULFATE ION, N-(3-tert-butyl-1H-pyrazol-5-yl)-2-[(2R)-2-methylpiperazin-1-yl]quinazolin-4-amine, ... (5 entities in total)
• Functional Keywords: aaa+ protein, atpase, hydrolase, enzyme-inhibitor complex, isomerase-inhibitor complex, isomerase/inhibitor
• Biological source: Drosophila melanogaster (Fruit fly)
• Total number of polymer chains: 1
• Total formula weight: 34900.74

Authors

Pisa, R.,Cupido, T.,Kapoor, T.M. (deposition date: 2019-02-12, release date: 2019-03-27, Last modification date: 2023-10-11)

Primary citation

Pisa, R.,Cupido, T.,Kapoor, T.M.
Designing Allele-Specific Inhibitors of Spastin, a Microtubule-Severing AAA Protein.
J. Am. Chem. Soc., 141:5602-5606, 2019

PubMed Abstract: The bump-hole approach is a powerful chemical biology strategy to specifically probe the functions of closely related proteins. However, for many protein families, such as the ATPases associated with diverse cellular activities (AAA), we lack structural data for inhibitor-protein complexes to design allele-specific chemical probes. Here we report the X-ray structure of a pyrazolylaminoquinazoline-based inhibitor bound to spastin, a microtubule-severing AAA protein, and characterize the residues involved in inhibitor binding. We show that an inhibitor analogue with a single-atom hydrogen-to-fluorine modification can selectively target a spastin allele with an engineered cysteine mutation in its active site. We also report an X-ray structure of the fluoro analogue bound to the spastin mutant. Furthermore, analyses of other mutant alleles suggest how the stereoelectronics of the fluorine-cysteine interaction, rather than sterics alone, contribute to the inhibitor-allele selectivity. This approach could be used to design allele-specific probes for studying cellular functions of spastin isoforms. Our data also suggest how tuning stereoelectronics can lead to specific inhibitor-allele pairs for the AAA superfamily.

PubMed: 30875216
DOI: 10.1021/jacs.8b13257

Experimental method

X-RAY DIFFRACTION (2.425 Å)