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. 2020 Jun 18;10:9940. doi: 10.1038/s41598-020-66852-7

Figure 2.

Figure 2

Solubilization of recombinant proteins from E. coli crude membranes. Membranes were incubated in 500 mM NaCl and 50 mM Tris/HCl, with 10% glycerol, pH 7.5, and 1% (w/v) of copolymer, and the samples were incubated for 2 h at RT for HR or at 37 °C for MsbA. For the detergent experiments, DDM was used at 1.5% for HR and a mixture of 2% DDM and 0.04% sodium cholate was used for MsbA. (A) Solubilization of HR. Det (n = 21): detergent; SMA (n = 20): 2:1 SMA from Malvern; zSMA: our synthetic zwitterionic copolymers that include 1:1 zSMA (n = 21) and 2:1 zSMA (n = 26) derived from the RAFT P(St-ran-MA) with molecular weights of 6.7 and 6.4 kDa, respectively; M zSMA: zwitterionic copolymers synthesized from Malvern SMA precursors that include 1:1 M zSMA (n = 15) and 2:1 M zSMA (n = 23) derived from the Malvern P(St-ran-MA) with molecular weights of 4.6 and 5.0 kDa, respectively. The white and red colors indicate 1:1 and 2:1 St:MA, respectively. Det solubilization was significantly higher than that with all polymers (P < 0.001). *Denotes P < 0.001 vs other copolymers; † denotes P < 0.02 vs 2:1 zSMA and 2:1 M zSMA. (B) Solubilization of MsbA. Det: n = 21; SMA: n = 20; 1:1 zSMA: n = 21; 2:1 zSMA: n = 25; 1:1 M zSMA: n = 15; and 2:1 M zSMA: n = 23. Det solubilization was significantly lower than that with all polymers (P < 0.001). *Denotes P < 0.01 vs other copolymers, except for 1:1 M zSMA (NS); †Denotes P < 0.001 vs corresponding M zSMAs; ‡Denotes P<0.001 vs 1:1 zSMA. Data are means ± SEM.