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A biosensor-based approach toward purification and crystallization of G protein-coupled receptors by Iva Navratilova; Marie Pancera; Richard T. Wyatt; David G. Myszka is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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Biacore technology was used to develop an aYnity puriWcation method and screen cocrystallization conditions for the chemokine receptor CCR5. We characterized the binding of nine HIV gp120 variants and identiWed a truncated construct (YU2DV1V2) that bound CCR5 independent of CD4. This construct was used in an aYnity puriWcation step to improve the activity of detergent-solubilized receptor by approximately 300%. The biosensor was also used to screen receptor binding activity automatically under 50 diVerent crystallization conditions. We found that high-molecular-weight polyethylene glycols (PEGs 4000 and 8000 Da) most often stabilized the receptor and improved complex formation with potential cocrystallization partners such as conformationally sensitive monoclonal antibodies and gp120. Our results show how biosensors can provide unique insights into receptor puriWcation methods and reveal the eVects of crystallization conditions on complex formation. Importantly, these methods can be readily applied to other systems.

Who reads A biosensor-based approach toward purification and crystallization of G protein-coupled receptors?

Author

Iva Navratilova; Marie Pancera; Richard T. Wyatt; David G. Myszka

Publisher

Elsevier Science; Elsevier ; Elsevier Inc.; Elsevier BV (ISSN 0003-2697)

Published

2006

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)