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High‐Throughput Detergent Exchange Screening Method for Membrane Protein Crystallization
Author(s) -
Lee Jonas,
Kim SungHou
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.2_supplement.280
Subject(s) - chemistry , elution , ion chromatography , transmembrane protein , chromatography , membrane protein , protein purification , membrane , ion exchange , affinity chromatography , crystallization , biochemistry , receptor , ion , organic chemistry , enzyme
We have coupled the immobilized metal ion affinity chromatography and desalting chromatography to exchange detergents of purified membrane proteins. Our method expedites the detergent exchange process by capturing the target with an affinity or ion‐exchange column, washing with a different detergent, and elute and desalt to remove the unwanted eluent from the target. We have coupled this method with the AkTA purification system to automate the detergent exchange of a purified membrane protein into six various detergents. This method was applied to screen several membrane protein targets for the crystallization trial including two transmembrane helices bacterial chemotaxis receptors and three transmembrane helices ExbB proton channel family protein. By applying this method, we were able to successfully crystallize E. coli ExbB protein concluding that this method could be applied as an efficient high‐throughput method to exchange and screen multiple detergents for membrane protein crystallization screening in the structural genomics platform. This work was supported by a National Institute Health Grant GM 62412.