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Automated three-dimensional reconstruction of keyhole limpet hemocyanin type 1

Academic Article
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Overview

authors

  • Mouche, F.
  • Zhu, Y. X.
  • Pulokas, J.
  • Potter, Clinton
  • Carragher, Bridget

publication date

  • 2003

journal

  • Journal of Structural Biology  Journal

abstract

  • We have reconstructed a three-dimensional map of keyhole limpet hemocyanin isoform 1 (KLH1), using our automated data collection software, Leginon, integrated with particle selection algorithms, and the SPIDER reconstruction package. KLH1, a 7.9 MDa macromolecule, is an extracellular respiratory pigment composed of two asymmetric decamers, and presents an overall D(5) point-group symmetry. The reconstruction is in agreement with previous data published on molluscan hemocyanins. The reconstructed map (11.3A resolution, 3sigma criterion) was used to fit an available X-ray crystallography structure of Octopus dofleini Odg, solved at 2.3A [J. Mol. Biol. 278 (4) (1998) 855], with satisfactory results. The results validate the approach of automating the cryoEM process and demonstrate that the quality of the images acquired and the particles selected is comparable to those obtained using manual methods. Several problems remain to be solved however before these results can be generalized.

subject areas

  • Algorithms
  • Animals
  • Calibration
  • Cryoelectron Microscopy
  • Crystallography, X-Ray
  • Databases as Topic
  • Dimerization
  • Hemocyanin
  • Image Processing, Computer-Assisted
  • Octopodiformes
  • Protein Conformation
  • Protein Isoforms
  • Software
  • Time Factors
  • X-Ray Diffraction
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Research

keywords

  • automated data acquisition
  • cryoelectron microscopy
  • hemocyanin
  • three-dimensional reconstruction
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Identity

International Standard Serial Number (ISSN)

  • 1047-8477

Digital Object Identifier (DOI)

  • 10.1016/j.jsb.2003.09.001

PubMed ID

  • 14643198
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Additional Document Info

start page

  • 301

end page

  • 312

volume

  • 144

issue

  • 3

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