Evidence is presented that two circumferential zones of cell envelope differentiation, the periseptal annuli, exist in E. coli as previously observed in S. typhimurium. The periseptal annulus is located at the division site of cells. A strain overproducing a periplasmic protein, PhoS (phosphate‐binding protein) has been used to provide a landmark for the periseptal compartment. The zone of adhesion does not involve inner‐outer membrane fusion. This zone does not provide a strong physical barrier to protein diffusion in the periplasmic space, at least under conditions of plasmolysis.