Crystallization of Nuclear Export Signals or Small-Molecule Inhibitors Bound to Nuclear Exporter CRM1.

Active Transport, Cell Nucleus Cell Nucleus / metabolism Crystallization / methods Crystallography, X-Ray / methods Karyopherins / chemistry Nuclear Export Signals Protein Binding Receptors, Cytoplasmic and Nuclear / chemistry Exportin 1 Protein
CRM1 KPT LMB Leptomycin B NES Nuclear export signals SINE X-ray crystallography XPO1

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2022
Historique:
entrez: 12 4 2022
pubmed: 13 4 2022
medline: 15 4 2022
Statut: ppublish

Résumé

The Karyopherin protein CRM1 or XPO1 is the major nuclear export receptor that regulates nuclear exit of thousands of macromolecules in the cell. CRM1 recognizes protein cargoes by binding to their 8-15 residue-long nuclear export signals (NESs). A ternary CRM1-Ran-RanBP1 complex engineered to be suitable for crystallization has enabled structure determination by X-ray crystallography of CRM1 bound to many NES peptides and small-molecule inhibitors. Here, we present a protocol for the purification of the individual proteins, formation of the ternary CRM1-Ran-RanBP1 complex and crystallization of this complex for X-ray crystallography.

Identifiants

DOI: 10.1007/978-1-0716-2337-4_19 PMID: 35412246
pubmed: 35412246
doi: 10.1007/978-1-0716-2337-4_19
doi:

Substances chimiques

Karyopherins 0
Nuclear Export Signals 0
Receptors, Cytoplasmic and Nuclear 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't Research Support, N.I.H., Extramural

Langues

eng

Sous-ensembles de citation

IM

Pagination

285-297

Subventions

Organisme : NIGMS NIH HHS
ID : R01 GM069909
Pays : United States

Informations de copyright

© 2022. Springer Science+Business Media, LLC, part of Springer Nature.

Références

Kirli K, Karaca S, Dehne HJ, Samwer M, Pan KT, Lenz C, Urlaub H, Gorlich D (2015) A deep proteomics perspective on CRM1-mediated nuclear export and nucleocytoplasmic partitioning. elife 4:e11466. https://doi.org/10.7554/eLife.11466
doi: 10.7554/eLife.11466 pubmed: 26673895 pmcid: 4764573
Koyama M, Matsuura Y (2010) An allosteric mechanism to displace nuclear export cargo from CRM1 and RanGTP by RanBP1. EMBO J 29(12):2002–2013. https://doi.org/10.1038/emboj.2010.89
doi: 10.1038/emboj.2010.89 pubmed: 20485264 pmcid: 2892370
Sun Q, Carrasco YP, Hu Y, Guo X, Mirzaei H, Macmillan J, Chook YM (2013) Nuclear export inhibition through covalent conjugation and hydrolysis of Leptomycin B by CRM1. Proc Natl Acad Sci U S A 110(4):1303–1308. https://doi.org/10.1073/pnas.1217203110
doi: 10.1073/pnas.1217203110 pubmed: 23297231 pmcid: 3557022
Lapalombella R, Sun Q, Williams K, Tangeman L, Jha S, Zhong Y, Goettl V, Mahoney E, Berglund C, Gupta S, Farmer A, Mani R, Johnson AJ, Lucas D, Mo X, Daelemans D, Sandanayaka V, Shechter S, McCauley D, Shacham S, Kauffman M, Chook YM, Byrd JC (2012) Selective inhibitors of nuclear export show that CRM1/XPO1 is a target in chronic lymphocytic leukemia. Blood 120(23):4621–4634. https://doi.org/10.1182/blood-2012-05-429506
doi: 10.1182/blood-2012-05-429506 pubmed: 23034282 pmcid: 3512237
Hing ZA, Fung HY, Ranganathan P, Mitchell S, El-Gamal D, Woyach JA, Williams K, Goettl VM, Smith J, Yu X, Meng X, Sun Q, Cagatay T, Lehman AM, Lucas DM, Baloglu E, Shacham S, Kauffman MG, Byrd JC, Chook YM, Garzon R, Lapalombella R (2016) Next-generation XPO1 inhibitor shows improved efficacy and in vivo tolerability in hematological malignancies. Leukemia 30(12):2364–2372. https://doi.org/10.1038/leu.2016.136
doi: 10.1038/leu.2016.136 pubmed: 27323910 pmcid: 5143172
Haines JD, Herbin O, de la Hera B, Vidaurre OG, Moy GA, Sun Q, Fung HY, Albrecht S, Alexandropoulos K, McCauley D, Chook YM, Kuhlmann T, Kidd GJ, Shacham S, Casaccia P (2015) Nuclear export inhibitors avert progression in preclinical models of inflammatory demyelination. Nat Neurosci 18(4):511–520. https://doi.org/10.1038/nn.3953
doi: 10.1038/nn.3953 pubmed: 25706475 pmcid: 4522902
Etchin J, Sun Q, Kentsis A, Farmer A, Zhang ZC, Sanda T, Mansour MR, Barcelo C, McCauley D, Kauffman M, Shacham S, Christie AL, Kung AL, Rodig SJ, Chook YM, Look AT (2013) Antileukemic activity of nuclear export inhibitors that spare normal hematopoietic cells. Leukemia 27(1):66–74. https://doi.org/10.1038/leu.2012.219
doi: 10.1038/leu.2012.219 pubmed: 22847027
Fung HY, Fu SC, Chook YM (2017) Nuclear export receptor CRM1 recognizes diverse conformations in nuclear export signals. elife 6:e23961. https://doi.org/10.7554/eLife.23961
doi: 10.7554/eLife.23961 pubmed: 28282025 pmcid: 5358978
Fung HY, Fu SC, Brautigam CA, Chook YM (2015) Structural determinants of nuclear export signal orientation in binding to exportin CRM1. elife 4:e10034. https://doi.org/10.7554/eLife.10034
doi: 10.7554/eLife.10034 pmcid: 4596688
Baumhardt JM, Walker JS, Lee Y, Shakya B, Brautigam CA, Lapalombella R, Grishin N, Chook YM (2020) Recognition of nuclear export signals by CRM1 carrying the oncogenic E571K mutation. Mol Biol Cell 31(17):1879–1891. https://doi.org/10.1091/mbc.E20-04-0233
doi: 10.1091/mbc.E20-04-0233 pubmed: 32520643 pmcid: 7525811

Auteurs

Ho Yee Joyce Fung (HYJ)

Department of Pharmacology, UT Southwestern Medical Center, Dallas, TX, USA.

Yuh Min Chook (YM)

Department of Pharmacology, UT Southwestern Medical Center, Dallas, TX, USA. yuhmin.chook@utsouthwestern.edu.

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Classifications MeSH

questionsmedicales.fr › Métabolisme › Transport biologique › Transport des protéines › Transport nucléaire actif › Crystallization of Nuclear Export Signals or...
questionsmedicales.fr › Cellules › Structures cellulaires › Espace intracellulaire › Cytoplasme › Structures cytoplasmiques › Organites › Noyau de la cellule › Crystallization of Nuclear Export Signals or...
questionsmedicales.fr › Phénomènes chimiques › Cristallisation › Crystallization of Nuclear Export Signals or...
questionsmedicales.fr › Techniques d'investigation › Techniques de chimie analytique › Cristallographie aux rayons X › Crystallization of Nuclear Export Signals or...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines nucléaires › Caryophérines › Crystallization of Nuclear Export Signals or...
questionsmedicales.fr › Phénomènes chimiques › Phénomènes biochimiques › Signaux de triage des protéines › Signaux d'export nucléaire › Crystallization of Nuclear Export Signals or...
questionsmedicales.fr › Métabolisme › Liaison aux protéines › Crystallization of Nuclear Export Signals or...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Récepteurs cytoplasmiques et nucléaires › Crystallization of Nuclear Export Signals or...