Open AccessManipulating phospholipids for crystallization of a membrane transport protein
Author(s) -
Lan Guan,
И. Н. Смирнова,
Gill Verner,
Shushi Nagamori,
H. Ronald Kaback
Publication year - 2006
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.0510922103
Subject(s) - crystallization , lactose permease , crystallography , protein crystallization , membrane protein , membrane , tetragonal crystal system , escherichia coli , resolution (logic) , chemistry , materials science , biophysics , membrane transport protein , crystal structure , biology , biochemistry , computer science , gene , organic chemistry , artificial intelligence
Crystallization is a major bottleneck to obtaining x-ray structures of membrane proteins. By applying an established crystallization protocol for the lactose permease (LacY) ofEscherichia coli , a systematic study of the effect of phospholipids (PL) on crystallization of LacY was undertaken. We observe three different crystal forms that diffract to increasingly better resolution in a manner that correlates with the concentration of copurified PL. Consistently, progressive addition ofE. coli PL to delipidated LacY leads to different crystal forms. Tetragonal crystals are obtained with improved diffraction quality for a stable mutant by carefully adjusting PL content. Furthermore, crystals of good quality from wild-type LacY, a particularly difficult protein, were also obtained by using same approach. Thus, it is likely that manipulation of PL is a good strategy for predominantly hydrophobic membrane proteins like LacY.