MicroED structure of lipid-embedded mammalian mitochondrial voltage-dependent anion channel.
FIB/SEM
MicroED
bicelle crystallization
cryoEM
microcrystal electron diffraction
Journal
Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876
Informations de publication
Date de publication:
22 12 2020
22 12 2020
Historique:
pubmed:
10
12
2020
medline:
9
2
2021
entrez:
9
12
2020
Statut:
ppublish
Résumé
A structure of the murine voltage-dependent anion channel (VDAC) was determined by microcrystal electron diffraction (MicroED). Microcrystals of an essential mutant of VDAC grew in a viscous bicelle suspension, making it unsuitable for conventional X-ray crystallography. Thin, plate-like crystals were identified using scanning-electron microscopy (SEM). Crystals were milled into thin lamellae using a focused-ion beam (FIB). MicroED data were collected from three crystal lamellae and merged for completeness. The refined structure revealed unmodeled densities between protein monomers, indicative of lipids that likely mediate contacts between the proteins in the crystal. This body of work demonstrates the effectiveness of milling membrane protein microcrystals grown in viscous media using a focused ion beam for subsequent structure determination by MicroED. This approach is well suited for samples that are intractable by X-ray crystallography. To our knowledge, the presented structure is a previously undescribed mutant of the membrane protein VDAC, crystallized in a lipid bicelle matrix and solved by MicroED.
Identifiants
pubmed: 33293416
pii: 2020010117
doi: 10.1073/pnas.2020010117
pmc: PMC7768720
doi:
Substances chimiques
Lipids
0
Mitochondrial Proteins
0
Voltage-Dependent Anion Channels
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
32380-32385Subventions
Organisme : NIGMS NIH HHS
ID : P41 GM136508
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM135175
Pays : United States
Organisme : Howard Hughes Medical Institute
Pays : United States
Déclaration de conflit d'intérêts
The authors declare no competing interest.
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