| Structural highlights
Function
[SPAST_DROME] ATP-dependent microtubule severing protein. Stimulates microtubule minus-end depolymerization and poleward microtubule flux in the mitotic spindle. Regulates microtubule stability in the neuromuscular junction synapse.[1] [2] [3] [4] [5] [6] [7]
Publication Abstract from PubMed
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 AAA (ATPases Associated with diverse cellular Activities) superfamily, we lack the structural data needed for the design of 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 char-acterize the residues involved in inhibitor binding. We show that an inhibitor analog with a single atom hydro-gen-to-fluorine modification can selectively target a spastin allele with an engineered cysteine mutation in its active site. We also report a x-ray structure of the fluoro-analog bound to the spastin mutant, along with analyses of other alleles with mutations at this position, that re-veal how stereoelectronics of the fluorine-cysteine inter-action, rather than sterics alone, contribute to inhibitor-allele selectivity. This approach could be used to design allele-specific probes for studying cellular functions of spastin isoforms. Our data suggests how tuning stereoe-lectronics can lead to specific inhibitor-allele pairs for the AAA superfamily.
Designing Allele-Specific Inhibitors of Spastin, a Microtubule-Severing AAA Protein.,Pisa R, Cupido T, Kapoor TM J Am Chem Soc. 2019 Mar 15. doi: 10.1021/jacs.8b13257. PMID:30875216[8]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Trotta N, Orso G, Rossetto MG, Daga A, Broadie K. The hereditary spastic paraplegia gene, spastin, regulates microtubule stability to modulate synaptic structure and function. Curr Biol. 2004 Jul 13;14(13):1135-47. PMID:15242610 doi:https://dx.doi.org/10.1016/j.cub.2004.06.058
- ↑ Sherwood NT, Sun Q, Xue M, Zhang B, Zinn K. Drosophila spastin regulates synaptic microtubule networks and is required for normal motor function. PLoS Biol. 2004 Dec;2(12):e429. Epub 2004 Nov 30. PMID:15562320 doi:https://dx.doi.org/10.1371/journal.pbio.0020429
- ↑ Roll-Mecak A, Vale RD. The Drosophila homologue of the hereditary spastic paraplegia protein, spastin, severs and disassembles microtubules. Curr Biol. 2005 Apr 12;15(7):650-5. PMID:15823537 doi:https://dx.doi.org/S0960-9822(05)00170-3
- ↑ Orso G, Martinuzzi A, Rossetto MG, Sartori E, Feany M, Daga A. Disease-related phenotypes in a Drosophila model of hereditary spastic paraplegia are ameliorated by treatment with vinblastine. J Clin Invest. 2005 Nov;115(11):3026-34. PMID:16276413 doi:https://dx.doi.org/10.1172/JCI24694
- ↑ Zhang D, Rogers GC, Buster DW, Sharp DJ. Three microtubule severing enzymes contribute to the "Pacman-flux" machinery that moves chromosomes. J Cell Biol. 2007 Apr 23;177(2):231-42. PMID:17452528 doi:https://dx.doi.org/jcb.200612011
- ↑ Lee M, Paik SK, Lee MJ, Kim YJ, Kim S, Nahm M, Oh SJ, Kim HM, Yim J, Lee CJ, Bae YC, Lee S. Drosophila Atlastin regulates the stability of muscle microtubules and is required for synapse development. Dev Biol. 2009 Jun 15;330(2):250-62. doi: 10.1016/j.ydbio.2009.03.019. Epub 2009 , Mar 31. PMID:19341724 doi:https://dx.doi.org/10.1016/j.ydbio.2009.03.019
- ↑ Roll-Mecak A, Vale RD. Structural basis of microtubule severing by the hereditary spastic paraplegia protein spastin. Nature. 2008 Jan 17;451(7176):363-7. PMID:18202664 doi:10.1038/nature06482
- ↑ Pisa R, Cupido T, Kapoor TM. Designing Allele-Specific Inhibitors of Spastin, a Microtubule-Severing AAA Protein. J Am Chem Soc. 2019 Mar 15. doi: 10.1021/jacs.8b13257. PMID:30875216 doi:https://dx.doi.org/10.1021/jacs.8b13257
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