title: Absence of long-range diffusion of OmpA in E. coli is not caused by its peptidoglycan binding domain.
abstract: BACKGROUND
  It is widely believed that integral outer membrane (OM) proteins in bacteria are able to diffuse laterally in the OM. However, stable, immobile proteins have been identified in the OM of Escherichia coli. In explaining the observations, a hypothesized interaction of the immobilized OM proteins with the underlying peptidoglycan (PG) cell wall played a prominent role.
  
  
  RESULTS
  OmpA is an abundant outer membrane protein in E. coli containing a PG-binding domain. We use FRAP to investigate whether OmpA is able to diffuse laterally over long-range (> ~100 nm) distances in the OM. First, we show that OmpA, containing a PG binding domain, does not exhibit long-range lateral diffusion in the OM. Then, to test whether PG interaction was required for this immobilization, we genetically removed the PG binding domain and repeated the FRAP experiment. To our surprise, this did not increase the mobility of the protein in the OM.
  
  
  CONCLUSIONS
  OmpA exhibits an absence of long-range (> ~100 nm) diffusion in the OM that is not caused by its PG binding domain. Therefore, other mechanisms are needed to explain this observation, such as the presence of physical barriers in the OM, or strong interactions with other elements in the cell envelope.
authors.forename: Gertjan S
authors.initials: GS
authors.lastname: Verhoeven
authors.forename: Marileen
authors.initials: M
authors.lastname: Dogterom
authors.forename: Tanneke
authors.initials: T
authors.lastname: den Blaauwen
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D001665:Binding Sites; D002462:Cell Membrane / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D036681:Fluorescence Recovery After Photobleaching; D010457:Peptidoglycan / Q000737:chemistry*; D021381:Protein Transport
chemical_list: D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D010457:Peptidoglycan; C078167:OMPA outer membrane proteins