Figure 3.

Hexameric particles isolated using size‐exclusion chromatography and single particle averaging.

A. Elution profile from size‐exclusion chromatography of His₆‐CotYc after nickel‐affinity purification with three major peaks labelled 1, 2 and 3 representing about 50%, 16% and 6% of the total eluted protein, respectively.

B. SDS‐PAGE of eluted fractions from size‐exclusion chromatography.

C. Negatively stained image of monodisperse His₆‐CotYc hexameric particles from fraction 3 (panel A). Scale bar represents 100 nm. Particles were obtained after protein purification from two independent cell batches.

D. Eigenimages from MSA; [1] is the rotational average of the particles; [2] shows the diameter variation; [3–5] indicate sixfold symmetry.

E. His₆‐CotYc class averages. Scale bar represents 10 nm.