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

Acta Crystallogr D Biol Crystallogr. 2004 Oct;60(Pt 10):1795-807. doi: 10.1107/S0907444904019109. Epub 2004 Sep 23.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Automation
  • Computers
  • Crystallization / methods*
  • Crystallography, X-Ray / instrumentation*
  • Crystallography, X-Ray / methods*
  • Image Processing, Computer-Assisted
  • Lipids / chemistry*
  • Membrane Proteins / chemistry*
  • Robotics
  • Software

Substances

  • Lipids
  • Membrane Proteins