title: Evaluation of SDS-polyacrylamide gel systems for the study of outer membrane protein profiles of clinical strains of Acinetobacter baumannii.
abstract: The outer membrane protein (OMP) profiles of 23 blood isolates of Acinetobacter baumannii representing all the different antimicrobial susceptibility patterns observed during a 3-year period in a Spanish hospital were studied. OMPs extracted from envelopes of sonicated cells after solubilisation with 2% of N-lauryl-sarcosinate were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) using the Laemmli's buffers. Eight running gel systems differing in the concentration of polyacrylamide (8%, 10% and 12%) and in the absence or presence of urea (4 M and 6 M) were used in a preliminary study analysing the OMP profiles of four clonally unrelated strains of A. baumannii. When this study was completed, the OMPs of the 23 A. baumannii were analysed in 10% SDS-polyacrylamide gels with 6 M urea and in 12% SDS-polyacrylamide gels. Ten OMP profiles were observed in 10% SDS-polyacrylamide gels with 6 M urea, whereas only 5 OMP profiles were visualised using 12% SDS-polyacrylamide gels. The OMP profiles obtained in 10% SDS-polyacrylamide gels with 6 M urea only partially correlated with those observed in 12% SDS-polyacrylamide gels. In conclusion, the use of 10% SDS-polyacrylamide gels with 6 M urea is recommended for the study of OMP profiles of A. baumannii.
authors.affiliation: Department of Microbiology, University of Seville, Apdo. 914, Spain. felipefc@supercable.es
authors.forename: Felipe Fernández
authors.initials: FF
authors.lastname: Cuenca
authors.forename: Alvaro
authors.initials: A
authors.lastname: Pascual
authors.forename: Luis
authors.initials: L
authors.lastname: Martínez Marínez
authors.forename: Ma Carmen
authors.initials: MC
authors.lastname: Conejo
authors.forename: Evelio J
authors.initials: EJ
authors.lastname: Perea
mesh_terms: D000151:Acinetobacter Infections / Q000453:epidemiology / Q000382:microbiology; D040981:Acinetobacter baumannii / Q000235:genetics / Q000302:isolation & purification*; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D003902:Detergents / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008970:Molecular Weight
chemical_list: D001425:Bacterial Outer Membrane Proteins; D003902:Detergents