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Titolo:
A strategy for determining the orientations of refractory particles for reconstruction from cryo-electron micrographs with particular reference to round, smooth-surfaced, icosahedral viruses
Autore:
Caston, JR; Belnap, DM; Steven, AC; Trus, BL;
Indirizzi:
NIAMSD, Struct Biol Res Lab, NIH, Bethesda, MD 20892 USA NIAMSD Bethesda MD USA 20892 ct Biol Res Lab, NIH, Bethesda, MD 20892 USA NIH,SAomputat Biosci & Engn Lab, Ctr Informat Technol, Bethesda, MD 20892 U NIH Bethesda MD USA 20892 Lab, Ctr Informat Technol, Bethesda, MD 20892 U
Titolo Testata:
JOURNAL OF STRUCTURAL BIOLOGY
fascicolo: 2-3, volume: 125, anno: 1999,
pagine: 209 - 215
SICI:
1047-8477(199904/05)125:2-3<209:ASFDTO>2.0.ZU;2-1
Fonte:
ISI
Lingua:
ENG
Soggetto:
X-RAY CRYSTALLOGRAPHY; DOUBLE-STRANDED-RNA; 3-DIMENSIONAL RECONSTRUCTION; CRYOELECTRON MICROSCOPY; BIOLOGICAL MACROMOLECULES; VISUALIZATION; CAPSIDS; CRYOMICROSCOPY; REPLICATION; RESOLUTION;
Keywords:
3D reconstruction; capsid structure; common lines;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
31
Recensione:
Indirizzi per estratti:
Indirizzo: Trus, BL NIAMSD, Struct Biol Res Lab, NIH, Bldg 12A,Room 2033,MSC 5624, Bethesda, MD NIAMSD Bldg 12A,Room 2033,MSC 5624 Bethesda MD USA 20892 esda, MD
Citazione:
J.R. Caston et al., "A strategy for determining the orientations of refractory particles for reconstruction from cryo-electron micrographs with particular reference to round, smooth-surfaced, icosahedral viruses", J STRUCT B, 125(2-3), 1999, pp. 209-215

Abstract

Cryo-electron microscopy and three-dimensional image reconstruction are powerful tools for analyzing icosahedral virus capsids at resolutions that now extend below 1 nm, However, the validity of such density maps depends critically on correct identification of the viewing geometry of each particle in the data set. In some cases-for example, round capsids with low surface relief-it is difficult to identify orientations by conventional applicationof the two most widely used approaches-"common lines" and model-based iterative refinement. We describe here a strategy for determining the orientations of such refractory specimens. The key step is to determine reliable orientations for a base set of particles, For each particle, a list of candidate orientations is generated by common lines: correct orientations are thenidentified by computing a single-particle reconstruction for each candidate and then systematically matching their reprojections with the original images by visual criteria and crosscorrelation analysis. This base set yieldsa first-generation reconstruction that is fed into the model-based procedure. This strategy has led to the structural determination of two viruses that, ill our hands, resisted solution by other means.

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Documento generato il 11/07/20 alle ore 20:42:21