Identification of aMoraxella catarrhalisOuter Membrane Protein Exhibiting Both Adhesin and Lipolytic Activities

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Abstract
The UspA1 and Hag proteins have previously been shown to be involved in the ability of theMoraxella catarrhaliswild-type strain O35E to bind to human Chang and A549 cells, respectively. In an effort to identify novel adhesins, we generated a plasmid library ofM. catarrhalisDNA fragments, which was introduced into a nonadherentEscherichia colistrain. RecombinantE. colibacteria 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 genemcaP(M. catarrhalisadherence protein). Sequence analysis revealed that McaP is related to autotransporter proteins and has substantial similarity with the GDSL family of lipolytic enzymes, particularly theMoraxella bovisphospholipase B. Expression of themcaPgene product byE. coliincreased adherence to Chang, A549, and 16HBE14o−polarized human bronchial cells 50- to 100-fold. Spectrophotometric assays withp-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 isogenicM. catarrhalisO35E mutants demonstrated that the lack of McaP expression abolishes esterase activity and considerably decreases adherence to several human cell lines.