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A robotic system for crystallizing membrane and soluble proteins in lipidic mesophases

Academic Article
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Overview

authors

  • Cherezov, Vadim
  • Peddi, A.
  • Muthusubramaniam, L.
  • Zheng, Y. F.
  • Caffrey, M.

publication date

  • October 2004

journal

  • Acta Crystallographica Section D-Biological Crystallography  Journal

abstract

  • A high-throughput robotic system has been developed for crystallizing membrane proteins using lipidic mesophases. It incorporates commercially available components and is relatively inexpensive. The crystallization robot uses standard automated liquid-handlers and a specially built device for accurately and reproducibly delivering nanolitre volumes of highly viscous protein/lipid mesophases. Under standard conditions, the robot uses just 20 nl protein solution, 30 nl lipid and 1 microl precipitant solution. 96 wells can be set up using the robot in 13 min. Trials are performed in specially designed 96-well glass plates. The slim (<2 mm high) plates have exquisite optical properties and are well suited for the detection of microcrystals and for birefringence-free imaging between crossed polarizers. Quantitative evaluation of the crystallization progress is performed using an automated imaging system. The optics, in combination with the slim crystallization plates, enables in-focus imaging of the entire well volume in a single shot such that a 96-well plate can be imaged in just 4.5 min. The performance characteristics of the robotic system and the versatility of the crystallization robot in performing vapor-diffusion, microbatch and bicelle crystallizations of membrane and soluble proteins are described.

subject areas

  • Automation
  • Computers
  • Crystallization
  • Crystallography, X-Ray
  • Image Processing, Computer-Assisted
  • Lipids
  • Membrane Proteins
  • Robotics
  • Software
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Identity

International Standard Serial Number (ISSN)

  • 0907-4449

Digital Object Identifier (DOI)

  • 10.1107/s0907444904019109

PubMed ID

  • 15388926
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Additional Document Info

start page

  • 1795

end page

  • 1807

volume

  • 60

issue

  • Pt 10

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