Figure 4: Extraction of L-series substrates in T-series liposomes.

A) Extraction of L20 in is most robust when there is a predicted hydrophobic mismatch. L20 substrate was reconstituted in liposomes T1-T3. Based on the length of the TMD and thickness of hydrophobic core of the lipid bilayer²⁴, there is a predicted hydrophobic mismatch T1, a possible mismatch in T2, and no predicted mismatch in T3.

B) Protease protection assay for proteoliposomes in A.

C) Similar to A, L22 substrate was reconstituted in liposomes T2-T4. There is a predicted hydrophobic mismatch T2, a possible mismatch in T3, and no predicted mismatch in T4.

D) Protease protection assay for proteoliposomes in C.

E) Similar to A, L24 substrate was reconstituted in liposomes T3 and T4. There is a predicted hydrophobic mismatch in T3 and no predicted mismatch in T4.

F) Protease protection assay for proteoliposomes in E.