Mannose-binding lectin binds to two major outer membrane proteins, opacity protein and porin, of Neisseria meningitidis.
Human mannose-binding lectin (MBL) provides a first line of defense against microorganisms by complement activation and/or opsonization in the absence of specific Ab. This serum collectin has been shown to activate complement when bound to repeating sugar moieties on several microorganisms, including encapsulated serogroup B and C meningococci, which leads to increased bacterial killing. In the present study, we sought to identify the meningococcal cell surface components to which MBL bound and to characterize such binding. Outer membrane complex containing both lipooligosaccharide (LOS) and proteins and LOS from Neisseria meningitidis were examined for MBL binding by dot blot and ELISA. MBL bound outer membrane complex but not LOS. The binding to bacteria by whole-cell ELISA did not require calcium and was not inhibited by N-acetyl-glucosamine or mannose. With the use of SDS-PAGE, immunoblot analysis, and mAbs specific for meningococcal opacity (Opa) proteins and porin proteins, we determined that MBL bound to Opa and porin protein B (porB). The N-terminal amino acid sequences of the two MBL binding proteins confirmed Opa and PorB. Purified PorB inhibited the binding of MBL to meningococci. Escherichia coli with surface-expressed gonococcal Opa bound significantly more MBL than did the control strain. The binding of human factor H to purified PorB was markedly inhibited by MBL in a dose-dependent manner. Meningococci incubated with human serum bound MBL as detected by ELISA. We conclude that MBL binds to meningococci by a novel target recognition of two nonglycosylated outer membrane proteins, Opa and PorB.
Estabrook
Michele M
MM
Center for Immunochemistry and Veterans Affairs Medical Center, San Francisco, CA 94121, USA. mme@itsa.ucsf.edu
Jack
Dominic L
DL
Klein
Nigel J
NJ
Jarvis
Gary A
GA
D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001426:Bacterial Proteins; D001667:Binding, Competitive / Q000276:immunology; D017242:Complement Factor H / Q000037:antagonists & inhibitors / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D037601:Mannose-Binding Lectin / Q000097:blood / Q000378:metabolism*; D009345:Neisseria meningitidis / Q000276:immunology*; D010446:Peptide Fragments / Q000378:metabolism; D018272:Porins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011485:Protein Binding / Q000276:immunology; D020539:Sequence Analysis, Protein
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; C493495:CFH protein, human; D037601:Mannose-Binding Lectin; D010446:Peptide Fragments; C419549:PorB protein, Chlamydia trachomatis; D018272:Porins; C057699:porin protein, Neisseria; C090348:Opa protein, Neisseria; D017242:Complement Factor H