Strategies for crystallizing membrane proteins.
Crystallizing membrane proteins remains a challenging endeavor despite the increasing number of membrane protein structures solved by X-ray crystallography. The critical factors in determining the success of the crystallization experiments are the purification and preparation of membrane protein samples. Moreover, there is the added complication that the crystallization conditions must be optimized for use in the presence of detergents although the methods used to crystallize most membrane proteins are, in essence, straightforward applications of standard methodologies for soluble protein crystallization. The roles that detergents play in stability and aggregation of membrane proteins as well as the colloidal properties of the protein-detergent complexes need to be appreciated and controlled before and during the crystallization trials. All X-ray quality crystals of membrane proteins were grown from preparations of detergent-solubilized protein, where the heterogeneous natural lipids from the membrane have been replaced by a homogeneous detergent environment. It is the preparation of such monodisperse, isotropic solutions of membrane proteins that has allowed the successful application of the standard crystallization methods routinely used on soluble proteins. In this review, the issues of protein purification and sample preparation are addressed as well as the new refinements in crystallization methodologies for membrane proteins. How the physical behavior of the detergent, in the form of micelles or protein-detergent aggregates, affects crystallization and the adaptation of published protocols to new membrane protein systems are also addressed. The general conclusion is that many integral membrane proteins could be crystallized if pure and monodisperse preparations in a suitable detergent system can be prepared.
Garavito
R M
RM
Department of Biochemistry and Molecular Biology, University Chicago, Illinois 60637, USA.
Picot
D
D
Loll
P J
PJ
D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D003902:Detergents; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification*; D018272:Porins / Q000737:chemistry / Q000302:isolation & purification; D011451:Prostaglandin-Endoperoxide Synthases / Q000737:chemistry / Q000302:isolation & purification; D013696:Temperature
D003902:Detergents; D008565:Membrane Proteins; D018272:Porins; D011451:Prostaglandin-Endoperoxide Synthases