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MSP1D1 and lipids preparation

WL Wu Liu
MS Martien Schoonen
TW Tong Wang
SM Sean McSweeney
QL Qun Liu
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Membrane scaffold protein MSP1D1 was expressed and purified from E. coli BL21 (DE3) pLysS cells40. Soybean polar lipids extracts (Avanti Polar Lipids, Inc., Birmingham, USA) were dissolved in chloroform and were transferred to glass tubes, dried under a nitrogen stream for 3 h. Residual chloroform was further removed by vacuum desiccation at 15 psi overnight, and the dried lipid films were resuspended in a buffer (100 mM sodium cholate, 20 mM HEPES, pH 7.6, 500 mM NaCl, 2 mM TCEP, 0.5 mM EDTA) and solicited for 1 hr in a bath sonicator until the liquid became clear. Finally, 50 mM soybean polar lipids were prepared in buffer containing 100 mM sodium cholate, 20 mM HEPES, pH 7.6, 500 mM NaCl, 2 mM TCEP, and 0.5 mM EDTA.

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