气候变化领域集成服务门户(CCPortal): CMT2N-causing aminoacylation domain mutants enable Nrp1 interaction with AlaRS

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DOI	10.1073/pnas.2012898118
	CMT2N-causing aminoacylation domain mutants enable Nrp1 interaction with AlaRS
	Sun L.; Wei N.; Kuhle B.; Blocquel D.; Novick S.; Matuszek Z.; Zhou H.; He W.; Zhang J.; Weber T.; Horvath R.; Latour P.; Pan T.; Schimmel P.; Griffin P.R.; Yang X.-L.
发表日期	2021
ISSN	00278424
卷号	118 期号:13
英文摘要	Through dominant mutations, aminoacyl-tRNA synthetases constitute the largest protein family linked to Charcot-Marie-Tooth disease (CMT). An example is CMT subtype 2N (CMT2N), caused by individual mutations spread out in AlaRS, including three in the aminoacylation domain, thereby suggesting a role for a tRNA-charging defect. However, here we found that two are aminoacylation defective but that the most widely distributed R329H is normal as a purified protein in vitro and in unfractionated patient cell samples. Remarkably, in contrast to wild-type (WT) AlaRS, all three mutant proteins gained the ability to interact with neuropilin 1 (Nrp1), the receptor previously linked to CMT pathogenesis in GlyRS. The aberrant AlaRS-Nrp1 interaction is further confirmed in patient samples carrying the R329H mutation. However, CMT2N mutations outside the aminoacylation domain do not induce the Nrp1 interaction. Detailed biochemical and biophysical investigations, including X-ray crystallography, small-angle X-ray scattering, hydrogen-deuterium exchange (HDX), switchSENSE hydrodynamic diameter determinations, and protease digestions reveal a mutation-induced structural loosening of the aminoacylation domain that correlates with the Nrp1 interaction. The b1b2 domains of Nrp1 are responsible for the interaction with R329H AlaRS. The results suggest Nrp1 is more broadly associated with CMT-associated members of the tRNA synthetase family. Moreover, we revealed a distinct structural loosening effect induced by a mutation in the editing domain and a lack of conformational impact with C-Ala domain mutations, indicating mutations in the same protein may cause neuropathy through different mechanisms. Our results show that, as with other CMT-associated tRNA synthetases, aminoacylation per se is not relevant to the pathology. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	AlaRS; Charcot-Marie-Tooth disease; Neuropilin 1
语种	英语
scopus关键词	amino acid transfer RNA ligase; neuropilin 1; protein AlaRS; protein CMT subtype 2N; proteinase; unclassified drug; Article; biochemical analysis; cell fractionation; clinical article; controlled study; deuterium hydrogen exchange; hereditary motor sensory neuropathy; human; human cell; hydrodynamics; in vitro study; pathogenesis; priority journal; protein conformation; protein protein interaction; protein purification; protein structure; tRNA aminoacylation; X ray crystallography
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/180098
作者单位	Department of Molecular Medicine, The Scripps Research Institute, San diego, CA 92037, United States; School of Public Health (Shenzhen), Sun Yat-sen University, Guangzhou, 510006, China; Department of Molecular Medicine, The Scripps Research Institute, Jupiter, FL 33458, United States; Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637, United States; School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, 510006, China; Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, 200237, China; Dynamic Biosensors GmbH, Martinsried, 82152, Germany; Department of Clinical Neurosciences, University of Cambridge, Cambridge, CB2 0PY, United Kingdom; Biology and Pathology Department, Hospices Civils, Lyon, 68500, France
推荐引用方式 GB/T 7714	Sun L.,Wei N.,Kuhle B.,et al. CMT2N-causing aminoacylation domain mutants enable Nrp1 interaction with AlaRS[J],2021,118(13).
APA	Sun L..,Wei N..,Kuhle B..,Blocquel D..,Novick S..,...&Yang X.-L..(2021).CMT2N-causing aminoacylation domain mutants enable Nrp1 interaction with AlaRS.Proceedings of the National Academy of Sciences of the United States of America,118(13).
MLA	Sun L.,et al."CMT2N-causing aminoacylation domain mutants enable Nrp1 interaction with AlaRS".Proceedings of the National Academy of Sciences of the United States of America 118.13(2021).