title: Detergents in Membrane Protein Purification and Crystallisation.
abstract: Detergents play a significant role in structural and functional characterisation of integral membrane proteins (IMPs). IMPs reside in the biological membranes and exhibit a great variation in their structural and physical properties. For in vitro biophysical studies, structural and functional analyses, IMPs need to be extracted from the membrane lipid bilayer environment in which they are found and purified to homogeneity while maintaining a folded and functionally active state. Detergents are capable of successfully solubilising and extracting the IMPs from the membrane bilayers. A number of detergents with varying structure and physicochemical properties are commercially available and can be applied for this purpose. Nevertheless, it is important to choose a detergent that is not only able to extract the membrane protein but also provide an optimal environment while retaining the correct structural and physical properties of the protein molecule. Choosing the best detergent for this task can be made possible by understanding the physical and chemical properties of the different detergents and their interaction with the IMPs. In addition, understanding the mechanism of membrane solubilisation and protein extraction along with crystallisation requirements, if crystallographic studies are going to be undertaken, can help in choosing the best detergent for the purpose. This chapter aims to present the fundamental properties of detergents and highlight information relevant to IMP crystallisation. The first section of the chapter reviews the physicochemical properties of detergents and parameters essential for predicting their behaviour in solution. The second section covers the interaction of detergents with the biologic membranes and proteins followed by their role in membrane protein crystallisation. The last section will briefly cover the types of detergent and their properties focusing on custom designed detergents for membrane protein studies.
authors.affiliation: School of Chemistry and Biochemistry, University of Western Australia, 35 Stirling Highway, Crawley, WA, 6009, Australia.
authors.forename: Anandhi
authors.initials: A
authors.lastname: Anandan
authors.affiliation: School of Chemistry and Biochemistry, University of Western Australia, 35 Stirling Highway, Crawley, WA, 6009, Australia. alice.vrielink@uwa.edu.au.
authors.forename: Alice
authors.initials: A
authors.lastname: Vrielink
mesh_terms: D000818:Animals; D055598:Chemical Phenomena; D003460:Crystallization; D018360:Crystallography, X-Ray; D003902:Detergents / Q000494:pharmacology*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008051:Lipid Bilayers; D008563:Membrane Lipids / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification*; D008823:Micelles; D008968:Molecular Conformation; D012995:Solubility
chemical_list: D003902:Detergents; D008051:Lipid Bilayers; D008563:Membrane Lipids; D008565:Membrane Proteins; D008823:Micelles
keywords: Crystallisation; Detergents; Lipids; Membrane proteins; Micelles; Protein purification