title: Quantification of detergent using colorimetric methods in membrane protein crystallography.
abstract: Membrane protein crystallography has the potential to greatly aid our understanding of membrane protein biology. Yet, membrane protein crystals remain challenging to produce. Although robust methods for the expression and purification of membrane proteins continue to be developed, the detergent component of membrane protein samples is equally important to crystallization efforts. This chapter describes the development of three colorimetric assays for the quantitation of detergent in membrane protein samples and provides detailed protocols. All of these techniques use small sample volumes and have potential applications in crystallography. The application of these techniques in crystallization prescreening, detergent concentration modification, and detergent exchange experiments is demonstrated. It has been observed that the concentration of detergent in a membrane protein sample can be just as important as the protein concentration when attempting to reproduce crystallization lead conditions.
authors.affiliation: Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, Canada.
authors.forename: Chelsy
authors.initials: C
authors.lastname: Prince
authors.affiliation: Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, Canada. Electronic address: jia@queensu.ca.
authors.forename: Zongchao
authors.initials: Z
authors.lastname: Jia
mesh_terms: D000818:Animals; D001647:Bile Acids and Salts / Q000737:chemistry; D003124:Colorimetry / Q000379:methods*; D003460:Crystallization / Q000379:methods*; D003902:Detergents / Q000032:analysis*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry*; D008982:Molybdenum / Q000737:chemistry; D013464:Sulfuric Acids / Q000737:chemistry; D014992:Xylenes / Q000737:chemistry
chemical_list: D001647:Bile Acids and Salts; D003902:Detergents; D008565:Membrane Proteins; D013464:Sulfuric Acids; D014992:Xylenes; C044659:molybdate; D008982:Molybdenum; C036531:2,6-xylenol; C033158:sulfuric acid
keywords: Analytical methods; Colorimetry; Crystallization; Detergent; Membrane protein; Quantification