title: Identification of a Moraxella catarrhalis outer membrane protein exhibiting both adhesin and lipolytic activities.
abstract: The UspA1 and Hag proteins have previously been shown to be involved in the ability of the Moraxella catarrhalis wild-type strain O35E to bind to human Chang and A549 cells, respectively. In an effort to identify novel adhesins, we generated a plasmid library of M. catarrhalis DNA fragments, which was introduced into a nonadherent Escherichia coli strain. Recombinant E. coli bacteria were subsequently enriched for clones that gained the ability to bind to Chang and A549 cells, yielding the plasmid pELFOS190. Transposon mutagenesis of this plasmid identified the potential adhesin gene mcaP (M. catarrhalis adherence protein). Sequence analysis revealed that McaP is related to autotransporter proteins and has substantial similarity with the GDSL family of lipolytic enzymes, particularly the Moraxella bovis phospholipase B. Expression of the mcaP gene product by E. coli increased adherence to Chang, A549, and 16HBE14o(-) polarized human bronchial cells 50- to 100-fold. Spectrophotometric assays with p-nitrophenol derivatives also demonstrated that McaP is an esterase. Furthermore, thin-layer chromatography revealed that McaP cleaves both phosphatidylcholine and lysophosphatidylcholine. McaP releases fatty acids and glycerophosphorylcholine upon cleavage of phosphatidylcholine, thus exhibiting phospholipase B activity. The construction and characterization of isogenic M. catarrhalis O35E mutants demonstrated that the lack of McaP expression abolishes esterase activity and considerably decreases adherence to several human cell lines.
authors.affiliation: Department of Microbiology and Immunology, Medical College of Ohio, Toledo, Ohio 43614-5806, USA.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Timpe
authors.forename: Melissa M
authors.initials: MM
authors.lastname: Holm
authors.forename: Serena L
authors.initials: SL
authors.lastname: Vanlerberg
authors.forename: Venkatesha
authors.initials: V
authors.lastname: Basrur
authors.forename: Eric R
authors.initials: ER
authors.lastname: Lafontaine
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000502:physiology*; D000595:Amino Acid Sequence; D001422:Bacterial Adhesion / Q000235:genetics / Q000502:physiology; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000502:physiology*; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D005798:Genes, Bacterial; D006801:Humans; D008066:Lipolysis / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D001936:Moraxella catarrhalis / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D016254:Mutagenesis, Insertional; D016366:Open Reading Frames; D010957:Plasmids / Q000235:genetics
chemical_list: D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial