3D variability analysis: Resolving continuous flexibility and discrete heterogeneity from single particle cryo-EM

[Display omitted] •Continuous structural heterogeneity of proteins is often functionally relevant.•Existing single particle cryo-EM reconstruction algorithms yield static structures.•Algorithm that fits linear subspace model at high resolution is introduced.•New method resolves detailed molecular fl...

Celý popis

Uložené v:
Podrobná bibliografia
Vydané v:Journal of structural biology Ročník 213; číslo 2; s. 107702
Hlavní autori: Punjani, Ali, Fleet, David J.
Médium: Journal Article
Jazyk:English
Vydavateľské údaje: United States Elsevier Inc 01.06.2021
Predmet:
Algorithms
Archaeal Proteins - chemistry
Continuous heterogeneity
Cryoelectron Microscopy - methods
Databases, Protein
Electron microscopy
Endopeptidases - chemistry
Expectation maximization algorithm
Image processing
Imaging, Three-Dimensional - methods
NAV1.7 Voltage-Gated Sodium Channel - chemistry
NAV1.7 Voltage-Gated Sodium Channel - metabolism
Plasmodium falciparum - chemistry
Principle component analysis
Receptors, Cannabinoid - chemistry
Ribosome Subunits, Large, Bacterial - chemistry
Ribosomes - chemistry
Signal-To-Noise Ratio
Single particle analysis
Spliceosomes - chemistry
Single particle analysis
Continuous heterogeneity
Image processing
Expectation maximization algorithm
Electron microscopy
Principle component analysis
ISSN:1047-8477, 1095-8657, 1095-8657
On-line prístup:Získať plný text
Tagy: Pridať tag
Žiadne tagy, Buďte prvý, kto otaguje tento záznam!
Popis
Shrnutí:[Display omitted] •Continuous structural heterogeneity of proteins is often functionally relevant.•Existing single particle cryo-EM reconstruction algorithms yield static structures.•Algorithm that fits linear subspace model at high resolution is introduced.•New method resolves detailed molecular flexibility from single particle data. Single particle cryo-EM excels in determining static structures of protein molecules, but existing 3D reconstruction methods have been ineffective in modelling flexible proteins. We introduce 3D variability analysis (3DVA), an algorithm that fits a linear subspace model of conformational change to cryo-EM data at high resolution. 3DVA enables the resolution and visualization of detailed molecular motions of both large and small proteins, revealing new biological insight from single particle cryo-EM data. Experimental results demonstrate the ability of 3DVA to resolve multiple flexible motions of α-helices in the sub-50 kDa transmembrane domain of a GPCR complex, bending modes of a sodium ion channel, five types of symmetric and symmetry-breaking flexibility in a proteasome, large motions in a spliceosome complex, and discrete conformational states of a ribosome assembly. 3DVA is implemented in the cryoSPARC software package.
Bibliografia:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1047-8477
1095-8657
1095-8657
DOI:10.1016/j.jsb.2021.107702