Journal: International Journal of Molecular Sciences
Article Title: Characterization of Adherent-Invasive Escherichia coli (AIEC) Outer Membrane Proteins Provides Potential Molecular Markers to Screen Putative AIEC Strains
doi: 10.3390/ijms23169005
Figure Lengend Snippet: OMP profiles of AIEC strains. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) OMP profiles of the AIEC strains grown in DMEM (2% bile salts) at 37 °C overnight: ( A ) 4I01, ( B ) 4C01, ( C ) 5I01, ( D ) 6I09, ( E ) 9C01, ( F ) 10I01, and ( G ) 18I08. Twelve percent polyacrylamide gels (13 cm; pH range: 4–7) were stained with Coomassie blue G-250. The scale bars on the left indicate molecular weights in kDa. Proteins with a well-known 2D-PAGE pattern are indicated (OmpA, OmpC, and Dps). ( H ) A representative AIEC OMP profile (blue spots) involving all the proteins present in at least 70% of the strains was superimposed on the commensal E. coli HS protein profile (orange spots). This analysis was performed using BioNumerics 2D software (v6.6). A total of 24 protein spots expressed differentially were selected and identified with MALDI-TOF/TOF mass spectrometry. These spots were absent in the HS OMP profile and highly frequent in the AIEC OMP profiles, as follows: 7/7 (100%, enclosed in green), 6/7 (86%, enclosed in purple), and 5/7 (71%, enclosed in red). * Spots 23 and 24 had no significant e-values in the MALDI-TOF/TOF analysis and were therefore discarded.
Article Snippet: Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) gels were analyzed using the software BioNumerics 2D (v6.6; Applied-Maths) as follows: The representative AIEC OMP 2D-PAGE profile was generated using the “ Matching gel 2D type ” tool, which can be used to combine gel images, using the molecular masses and isoelectric point (pI) values as a reference.
Techniques: Polyacrylamide Gel Electrophoresis, Staining, Software, Mass Spectrometry
