Structure Determination from Lipidic Cubic Phase Embedded Microcrystals by MicroED.

Cholesterol / chemistry Cryoelectron Microscopy / methods Crystallization / methods Endopeptidase K / chemistry Glycols / chemistry Lipids / chemistry Microscopy, Electron, Transmission / methods Nanoparticles / chemistry Receptor, Adenosine A2A / chemistry
GPCR Proteinase K additive phase conversion cholesterol cryo-electron microscopy (cryo-EM) lipase hydrolysis lipidic cubic phase (LCP) membrane protein microcrystal electron diffraction (MicroED) microcrystallography

Journal

Structure (London, England : 1993)
ISSN: 1878-4186
Titre abrégé: Structure
Pays: United States
ID NLM: 101087697

Informations de publication

Date de publication:
06 10 2020
Historique:
received: 02 03 2020
revised: 02 06 2020
accepted: 13 07 2020
pubmed: 1 8 2020
medline: 14 8 2021
entrez: 1 8 2020
Statut: ppublish

Résumé

The lipidic cubic phase (LCP) technique has proved to facilitate the growth of high-quality crystals that are otherwise difficult to grow by other methods. However, the crystal size optimization process could be time and resource consuming, if it ever happens. Therefore, improved techniques for structure determination using these small crystals is an important strategy in diffraction technology development. Microcrystal electron diffraction (MicroED) is a technique that uses a cryo-transmission electron microscopy to collect electron diffraction data and determine high-resolution structures from very thin micro- and nanocrystals. In this work, we have used modified LCP and MicroED protocols to analyze crystals embedded in LCP converted by 2-methyl-2,4-pentanediol or lipase, including Proteinase K crystals grown in solution, cholesterol crystals, and human adenosine A

Identifiants

DOI: 10.1016/j.str.2020.07.006 PMID: 32735770 PMC: PMC7544639
pubmed: 32735770
pii: S0969-2126(20)30239-2
doi: 10.1016/j.str.2020.07.006
pmc: PMC7544639
mid: NIHMS1617055
pii:
doi:

Substances chimiques

ADORA2A protein, human 0
Glycols 0
Lipids 0
Receptor, Adenosine A2A 0
Cholesterol 97C5T2UQ7J
Endopeptidase K EC 3.4.21.64
hexylene glycol KEH0A3F75J

Types de publication

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

Langues

eng

Sous-ensembles de citation

IM

Pagination

1149-1159.e4

Subventions

Organisme : NIGMS NIH HHS
ID : P41 GM136508
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM124152
Pays : United States
Organisme : NIDA NIH HHS
ID : R21 DA042298
Pays : United States
Organisme : Howard Hughes Medical Institute
Pays : United States

Commentaires et corrections

Type : CommentIn

Informations de copyright

Copyright © 2020 Elsevier Ltd. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of Interests The authors declare no competing interests.

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Auteurs

Lan Zhu (L)

Biodesign Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA; School of Molecular Sciences, Arizona State University, 551 East University Drive, Tempe, AZ 85287, USA.

Guanhong Bu (G)

Biodesign Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA; School for Engineering of Matter, Transport and Energy, Arizona State University, Tempe, AZ, USA.

Liang Jing (L)

Biodesign Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA; School of Molecular Sciences, Arizona State University, 551 East University Drive, Tempe, AZ 85287, USA.

Dan Shi (D)

Structural Biophysics Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD, USA.

Ming-Yue Lee (MY)

Biodesign Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA; School of Molecular Sciences, Arizona State University, 551 East University Drive, Tempe, AZ 85287, USA.

Tamir Gonen (T)

Howard Hughes Medical Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA; Departments of Biological Chemistry and Physiology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.

Wei Liu (W)

Biodesign Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA; School of Molecular Sciences, Arizona State University, 551 East University Drive, Tempe, AZ 85287, USA. Electronic address: w.liu@asu.edu.

Brent L Nannenga (BL)

Biodesign Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA; School for Engineering of Matter, Transport and Energy, Arizona State University, Tempe, AZ, USA. Electronic address: brent.nannenga@asu.edu.

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Classifications MeSH

questionsmedicales.fr › Lipides › Lipides membranaires › Stérols › Cholestérol › Structure Determination from Lipidic Cubic...
questionsmedicales.fr › Techniques d'investigation › Microscopie électronique › Cryomicroscopie électronique › Structure Determination from Lipidic Cubic...
questionsmedicales.fr › Phénomènes chimiques › Cristallisation › Structure Determination from Lipidic Cubic...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Hydrolases › Peptide hydrolases › Protéases à sérine › Serine endopeptidases › Endopeptidase K › Structure Determination from Lipidic Cubic...
questionsmedicales.fr › Composés chimiques organiques › Alcools › Glycols › Structure Determination from Lipidic Cubic...
questionsmedicales.fr › Lipides › Structure Determination from Lipidic Cubic...
questionsmedicales.fr › Techniques d'investigation › Microscopie électronique › Microscopie électronique à transmission › Structure Determination from Lipidic Cubic...
questionsmedicales.fr › Technologie, industrie et agriculture › Produits manufacturés › Nanostructures › Nanoparticules › Structure Determination from Lipidic Cubic...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines membranaires › Récepteurs de surface cellulaire › Récepteurs aux neuromédiateurs › Récepteurs purinergiques › Récepteurs purinergiques P1 › Récepteurs A2 à l'adénosine › Récepteur A2A à l'adénosine › Structure Determination from Lipidic Cubic...