A 11.5 Å single particle reconstruction of GroEL using EMAN

Steven J. Ludtke, Joanita Jakana, Jiu Li Song, David T. Chuang, Wah Chiu

Research output: Contribution to journalArticle

114 Citations (Scopus)

Abstract

Single-particle analysis has become an increasingly important method for structural determination of large macromolecular assemblies. GroEL is an 800 kDa molecular chaperone, which, along with its co-chaperonin GroES, promotes protein folding both in vitro and in the bacterial cell. EMAN is a single-particle analysis software package, which was first publicly distributed in 2000. We present a three-dimensional reconstruction of native naked GroEL to ∼11.5 Å performed entirely with EMAN. We demonstrate that the single-particle reconstruction, X-ray scattering data and X-ray crystal structure all agree well at this resolution. These results validate the specific methods of image restoration, reconstruction and evaluation techniques implemented in EMAN. It also demonstrates that the single-particle reconstruction technique and X-ray crystallography will yield consistent structure factors, even at low resolution, when image restoration is performed correctly. A detailed comparison of the single-particle and X-ray structures exhibits some small variations in the equatorial domain of the molecule, likely due to the absence of crystal packing forces in the single-particle reconstruction.

Original languageEnglish (US)
Pages (from-to)253-262
Number of pages10
JournalJournal of Molecular Biology
Volume314
Issue number2
StatePublished - Nov 23 2001

Fingerprint

X-Rays
Chaperonin 10
Chaperonins
Molecular Chaperones
Computer-Assisted Image Processing
X Ray Crystallography
Protein Folding
Software
In Vitro Techniques

Keywords

  • Electron cryomicroscopy
  • GroEL
  • Single-particle reconstruction
  • Structure
  • X-ray scattering

ASJC Scopus subject areas

  • Virology

Cite this

Ludtke, S. J., Jakana, J., Song, J. L., Chuang, D. T., & Chiu, W. (2001). A 11.5 Å single particle reconstruction of GroEL using EMAN. Journal of Molecular Biology, 314(2), 253-262.

A 11.5 Å single particle reconstruction of GroEL using EMAN. / Ludtke, Steven J.; Jakana, Joanita; Song, Jiu Li; Chuang, David T.; Chiu, Wah.

In: Journal of Molecular Biology, Vol. 314, No. 2, 23.11.2001, p. 253-262.

Research output: Contribution to journalArticle

Ludtke, SJ, Jakana, J, Song, JL, Chuang, DT & Chiu, W 2001, 'A 11.5 Å single particle reconstruction of GroEL using EMAN', Journal of Molecular Biology, vol. 314, no. 2, pp. 253-262.
Ludtke SJ, Jakana J, Song JL, Chuang DT, Chiu W. A 11.5 Å single particle reconstruction of GroEL using EMAN. Journal of Molecular Biology. 2001 Nov 23;314(2):253-262.
Ludtke, Steven J. ; Jakana, Joanita ; Song, Jiu Li ; Chuang, David T. ; Chiu, Wah. / A 11.5 Å single particle reconstruction of GroEL using EMAN. In: Journal of Molecular Biology. 2001 ; Vol. 314, No. 2. pp. 253-262.
@article{aca5817dc6094884b3a6f3c02e31d208,
title = "A 11.5 {\AA} single particle reconstruction of GroEL using EMAN",
abstract = "Single-particle analysis has become an increasingly important method for structural determination of large macromolecular assemblies. GroEL is an 800 kDa molecular chaperone, which, along with its co-chaperonin GroES, promotes protein folding both in vitro and in the bacterial cell. EMAN is a single-particle analysis software package, which was first publicly distributed in 2000. We present a three-dimensional reconstruction of native naked GroEL to ∼11.5 {\AA} performed entirely with EMAN. We demonstrate that the single-particle reconstruction, X-ray scattering data and X-ray crystal structure all agree well at this resolution. These results validate the specific methods of image restoration, reconstruction and evaluation techniques implemented in EMAN. It also demonstrates that the single-particle reconstruction technique and X-ray crystallography will yield consistent structure factors, even at low resolution, when image restoration is performed correctly. A detailed comparison of the single-particle and X-ray structures exhibits some small variations in the equatorial domain of the molecule, likely due to the absence of crystal packing forces in the single-particle reconstruction.",
keywords = "Electron cryomicroscopy, GroEL, Single-particle reconstruction, Structure, X-ray scattering",
author = "Ludtke, {Steven J.} and Joanita Jakana and Song, {Jiu Li} and Chuang, {David T.} and Wah Chiu",
year = "2001",
month = "11",
day = "23",
language = "English (US)",
volume = "314",
pages = "253--262",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - A 11.5 Å single particle reconstruction of GroEL using EMAN

AU - Ludtke, Steven J.

AU - Jakana, Joanita

AU - Song, Jiu Li

AU - Chuang, David T.

AU - Chiu, Wah

PY - 2001/11/23

Y1 - 2001/11/23

N2 - Single-particle analysis has become an increasingly important method for structural determination of large macromolecular assemblies. GroEL is an 800 kDa molecular chaperone, which, along with its co-chaperonin GroES, promotes protein folding both in vitro and in the bacterial cell. EMAN is a single-particle analysis software package, which was first publicly distributed in 2000. We present a three-dimensional reconstruction of native naked GroEL to ∼11.5 Å performed entirely with EMAN. We demonstrate that the single-particle reconstruction, X-ray scattering data and X-ray crystal structure all agree well at this resolution. These results validate the specific methods of image restoration, reconstruction and evaluation techniques implemented in EMAN. It also demonstrates that the single-particle reconstruction technique and X-ray crystallography will yield consistent structure factors, even at low resolution, when image restoration is performed correctly. A detailed comparison of the single-particle and X-ray structures exhibits some small variations in the equatorial domain of the molecule, likely due to the absence of crystal packing forces in the single-particle reconstruction.

AB - Single-particle analysis has become an increasingly important method for structural determination of large macromolecular assemblies. GroEL is an 800 kDa molecular chaperone, which, along with its co-chaperonin GroES, promotes protein folding both in vitro and in the bacterial cell. EMAN is a single-particle analysis software package, which was first publicly distributed in 2000. We present a three-dimensional reconstruction of native naked GroEL to ∼11.5 Å performed entirely with EMAN. We demonstrate that the single-particle reconstruction, X-ray scattering data and X-ray crystal structure all agree well at this resolution. These results validate the specific methods of image restoration, reconstruction and evaluation techniques implemented in EMAN. It also demonstrates that the single-particle reconstruction technique and X-ray crystallography will yield consistent structure factors, even at low resolution, when image restoration is performed correctly. A detailed comparison of the single-particle and X-ray structures exhibits some small variations in the equatorial domain of the molecule, likely due to the absence of crystal packing forces in the single-particle reconstruction.

KW - Electron cryomicroscopy

KW - GroEL

KW - Single-particle reconstruction

KW - Structure

KW - X-ray scattering

UR - http://www.scopus.com/inward/record.url?scp=0035940689&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035940689&partnerID=8YFLogxK

M3 - Article

VL - 314

SP - 253

EP - 262

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 2

ER -