Solving a new R2lox protein structure by microcrystal electron diffraction.


Journal

Science advances
ISSN: 2375-2548
Titre abrégé: Sci Adv
Pays: United States
ID NLM: 101653440

Informations de publication

Date de publication:
08 2019
Historique:
received: 25 03 2019
accepted: 27 06 2019
entrez: 29 8 2019
pubmed: 29 8 2019
medline: 6 5 2020
Statut: epublish

Résumé

Microcrystal electron diffraction (MicroED) has recently shown potential for structural biology. It enables the study of biomolecules from micrometer-sized 3D crystals that are too small to be studied by conventional x-ray crystallography. However, to date, MicroED has only been applied to redetermine protein structures that had already been solved previously by x-ray diffraction. Here, we present the first new protein structure-an R2lox enzyme-solved using MicroED. The structure was phased by molecular replacement using a search model of 35% sequence identity. The resulting electrostatic scattering potential map at 3.0-Å resolution was of sufficient quality to allow accurate model building and refinement. The dinuclear metal cofactor could be located in the map and was modeled as a heterodinuclear Mn/Fe center based on previous studies. Our results demonstrate that MicroED has the potential to become a widely applicable tool for revealing novel insights into protein structure and function.

Identifiants

pubmed: 31457106
doi: 10.1126/sciadv.aax4621
pii: aax4621
pmc: PMC6685719
doi:

Substances chimiques

Archaeal Proteins 0
Flavoproteins 0
Metalloproteins 0
Recombinant Proteins 0
metalloflavoproteins 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

eaax4621

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Auteurs

Hongyi Xu (H)

Department of Materials and Environmental Chemistry, Stockholm University, 10691 Stockholm, Sweden.

Hugo Lebrette (H)

Department of Biochemistry and Biophysics, Stockholm University, 10691 Stockholm, Sweden.

Max T B Clabbers (MTB)

Department of Materials and Environmental Chemistry, Stockholm University, 10691 Stockholm, Sweden.

Jingjing Zhao (J)

Department of Materials and Environmental Chemistry, Stockholm University, 10691 Stockholm, Sweden.

Julia J Griese (JJ)

Department of Biochemistry and Biophysics, Stockholm University, 10691 Stockholm, Sweden.
Department of Cell and Molecular Biology, Uppsala University, 75124 Uppsala, Sweden.

Xiaodong Zou (X)

Department of Materials and Environmental Chemistry, Stockholm University, 10691 Stockholm, Sweden.

Martin Högbom (M)

Department of Biochemistry and Biophysics, Stockholm University, 10691 Stockholm, Sweden.

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Classifications MeSH