Super-Resolution Fluorescence Microscopy Study ofthe Production of K1 Capsules by Escherichia coli: Evidence for the Differential Distribution of the Capsule at thePoles and the Equator of the Cell

摘要

The production of Escherichia coli K1 serotype capsule was investigated using direct stochastic optical reconstruction microscopy with live bacteria and graphene oxide-coated coverslips, overcoming many morphological artifacts found in other high-resolution imaging techniques. Super-resolution fluorescence images showed that the K1 capsular polysaccharide is not uniformly distributed on the cell surface, as previously thought. These studies demonstrated that on the cell surfaces the K1 capsule at the poles had bimodal thicknesses of 238 ± 41 and 323 ± 62 nm, whereas at the equator, there was a monomodal thickness of 217 ± 29 nm. This bimodal variation was also observed in high-pressure light-scattering chromatography measurements of purified K1 capsular polysaccharide. Particle tracking demonstrated that the formation of the capsule was dominated by the expansion of lyso-phosphatidylglycerol (lyso-PG) rafts that anchor the capsular polysaccharide in the outer membrane, and the expansion of these rafts across the cell surface was driven by new materialtransported through the capsular biosynthesis channels. The discoveryof thicker capsules at the poles of the cell will have implicationsin mediating interactions between the bacterium and its immediateenvironment.