High-throughput expression and detergent screening of integral membrane proteins.
Integral membrane proteins are a major challenge within structural genomics. These proteins are not only difficult to produce in quantities sufficient for analysis by X-ray diffraction or NMR, but also require extraction from their lipid environment, which leads to a new dimension of difficulties in purification and subsequent structural analysis. To overcome these problems requires new strategies enabling screening larger number of parameters dealing with expression and purification. For this reason, we have developed high-throughput methods for screening extracting and purifying detergents as well as other purification parameters, e.g. salt and pH. The method requires standard laboratory equipments, but can also be automated.
Eshaghi
Said
S
Division of Biophysics, Department of Medical Biochemistry and Biophysics, Karolinska Instiutet, Stockholm, Sweden.
D000818:Animals; D003902:Detergents / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification*; D012492:Salts / Q000737:chemistry
D003902:Detergents; D008565:Membrane Proteins; D011994:Recombinant Proteins; D012492:Salts