Skip to main content
. Author manuscript; available in PMC: 2021 Jul 22.
Published in final edited form as: J Am Chem Soc. 2020 Jul 8;142(29):12715–12729. doi: 10.1021/jacs.0c04669

Figure 7. C99 forms oligomers in SCOR bicelles.

Figure 7.

A) FRET spectra of IAEDANS-labeled C99 at a fixed protein to bicelle concentration ratio in the presence of increasing amounts of IANBD-labeled C99. The mole:mole ratios for each scan, in order of decreasing donor intensity, are (donor: acceptor): 1:0, 1:0.25, 1:1, 1:1.5, 1:2, and 1:3. B) Linear fit of normalized FRET efficiency from scans in A. Error bars represent the error in determination of the FRET transfer efficiency. C) DEER spectra of MTSL-labeled 200 μM C99 (L723C mutant) in 5 wt% SCOR bicelles, q=0.33. D) DEER spectra of MTSL-labeled 200 μM C99 (K699C mutant) in 5 wt % SCOR bicelles, q=0.33. E) Example SDS-PAGE (coomassie staining) of purified C99 following glutaraldehyde crosslinking of C99 in SCOR bicelles. Monomer, dimer, and trimer bands are indicated by M, D, and T respectively. F) Quantification of monomeric C99 from SDS-PAGE crosslinking experiments with C99, where the purification tag has been shortened and moved from the C to N terminus. Experiments were conducted with 50 μM C99 and various concentrations of SCOR bicelles in NMR buffer at 25 ° C n=3. The legend indicates the protein to lipid ratio.