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Titolo:
Spherical reconstruction: A method for structure determination of membraneproteins from cryo-EM images
Autore:
Jiang, QX; Chester, DW; Sigworth, FJ;
Indirizzi:
Yale Univ, Dept Cellular & Mol Physiol, New Haven, CT 06520 USA Yale UnivNew Haven CT USA 06520 r & Mol Physiol, New Haven, CT 06520 USA
Titolo Testata:
JOURNAL OF STRUCTURAL BIOLOGY
fascicolo: 2-3, volume: 133, anno: 2001,
pagine: 119 - 131
SICI:
1047-8477(200102/03)133:2-3<119:SRAMFS>2.0.ZU;2-U
Fonte:
ISI
Lingua:
ENG
Soggetto:
COLI 70S RIBOSOME; CRYOELECTRON MICROSCOPY; ESCHERICHIA-COLI; ELECTRON CRYOMICROSCOPY; 3-DIMENSIONAL STRUCTURE; VISUALIZATION; RESOLUTION; ANGSTROM; TRANSLOCATION; REFINEMENT;
Keywords:
spherical reconstruction; cryo-electron microscopy; membrane proteins; reconstitution; single-particle reconstruction; Euler angles;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
33
Recensione:
Indirizzi per estratti:
Indirizzo: Jiang, QX Yale Univ, Dept Cellular & Mol Physiol, 333 Cedar St, New Haven,CT 06520 USA Yale Univ 333 Cedar St New Haven CT USA 06520 aven, CT 06520 USA
Citazione:
Q.X. Jiang et al., "Spherical reconstruction: A method for structure determination of membraneproteins from cryo-EM images", J STRUCT B, 133(2-3), 2001, pp. 119-131

Abstract

We propose a new method for single-particle reconstruction, which should be generally applicable to structure determination for membrane proteins. After reconstitution into a small spherical vesicle, a membrane protein takesa particular orientation relative to the membrane normal, and its positionin the projected image of the vesicle directly defines two of its three Euler angles of orientation. The spherical constraint imposed by the vesicle effectively reduces the dimensionality of the alignment search from 5 to 3 and simplifies the detection of the particle. Projection images of particles in vesicles collectively take all possible orientations and therefore cover the whole Fourier space. Analysis of images of vesicles in ice showed that the vesicle density is well described by a simple model for membrane electron scattering density. In fitting this model we found that osmotically swollen vesicles remain nearly spherical through the freezing process. Theseresults satisfy the basic experimental requirements for spherical reconstruction. A computer simulation of particles in vesicles showed that this method provides good estimates of the two Euler angles and thus may improve single-particle reconstruction and extend it to smaller membrane proteins. (C) 2001 Academic Press.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 15/08/20 alle ore 20:00:10