by Peter Nollert
December 29, 2010 03:42
An anonymous commenter has supplied me with a reference, detailing the use of filter plates as standard protocol for the identification of useful membrane protein expression constructs.
Love J, Mancia F, Shapiro L, Punta M, Rost B, Girvin M, Wang DN, Zhou M, Hunt JF, Szyperski T, Gouaux E, MacKinnon R, McDermott A, Honig B, Inouye M, Montelione G, & Hendrickson WA (2010). The New York Consortium on Membrane Protein Structure (NYCOMPS): a high-throughput platform for structural genomics of integral membrane proteins. Journal of structural and functional genomics, 11 (3), 191-9 PMID: 20690043
The filter plate based method is related to the detergent scouting protocol described in
Vergis JM, Purdy MD, & Wiener MC (2010). A high-throughput differential filtration assay to screen and select detergents for membrane proteins. Analytical biochemistry, 407 (1), 1-11 PMID: 20667442
and reviewed in this recent blog post. There's a significant difference though: here only a single detergent, dodecyl maltoside, is applied while assaying the solubilization behavior of many different membrane protein expression constructs. The protein, after all is the most important crystallization parameter.
Cheers,
Peter