Figure 2.

Ability of apoA-IV^64-335 to model apoA-IV^WT. (A) Isolated apoA-IV^WT and apoA-IV^64-335 dimer incubated at 37°C in PBS for 24hrs and analyzed by SEC (dimers at 11–12 ml, monomers at 14–15 ml). (B) Far UV CD spectra of dimeric apoA-IV^WT and apoA-IV^64-335 at 0.85μM at 25°C. (C) Thermal denaturation of both forms as monitored by CD at 222nm. (D) Rate of dimer disassociation over time measured by SEC for both apoA-IV^WT and apoA-IV^64-335. (E) Introduction of single Cys point mutations (N151C, E263C, K233C and R284C) to test the dimer model. Predicted disulfide bond formation in WT apoA-IV based on the crystal structure is indicated with a solid line. (F) SDS-PAGE of point mutations of apoA-IV^WT designed in (D) that were purified under reducing conditions (right) and then oxidized to allow disulfide bond formation (left), stained with coomassie blue. See also Figure S2 and Table S1.