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Improving protein crystal quality by decoupling nucleation and growth in vapor diffusion

Published online by Cambridge University Press:  01 April 2000

EMMANUEL SARIDAKIS
Affiliation:
Division of Biomedical Sciences, Sir Alexander Fleming Building, Imperial College of Science, Technology and Medicine, London SW7 2AZ, United Kingdom
NAOMI E. CHAYEN
Affiliation:
Division of Biomedical Sciences, Sir Alexander Fleming Building, Imperial College of Science, Technology and Medicine, London SW7 2AZ, United Kingdom
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Abstract

A simple method for growing protein crystals in the metastable zone using the vapor diffusion technique is described. The coverslips holding the hanging drops are transferred, after being incubated for some time at conditions normally giving many small crystals, over reservoirs at concentrations that normally yield clear drops. Fewer, much larger and better diffracting crystals are obtained, compared with conventional crystallization at similar conditions. To our knowledge, this is the first report of a significant crystal improvement due to “backing off” from nucleation conditions, using the hanging drop method.

A correlation of the transfer time with published results for vapor diffusion equilibration of poly(ethylene glycol) solutions is also presented.

Type
Research Article
Copyright
© 2000 The Protein Society

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