Interaction between complement subcomponent C1q and the Klebsiella pneumoniae porin OmpK36.
The interaction between C1q, a subcomponent of the complement classical pathway component C1, and OmpK36, a porin protein from Klebsiella pneumoniae, was studied in a solid-phase direct-binding assay, inhibition assays with the purified globular and collagen-like regions of C1q, and cross-linking experiments. We have shown that the binding of C1q to the OmpK36 porin of the serum-sensitive strain K. pneumoniae KT707 occurs in an in vivo situation and that this binding leads to activation of the complement classical pathway and the subsequent deposition of complement components C3b and C5b-9 on the OmpK36 porin. Scatchard analysis of the binding of [125I]C1q to the OmpK36 porin showed two binding sites with dissociation constants of 1.5 and 75 nM. The decrease of [125I]C1q binding to the OmpK36 porin in buffer with increasing salt concentrations and the pIs of the C1q subcomponent (10.3) and OmpK36 porin (4.5) suggest that charged amino acids are involved in the binding phenomenon. In inhibition assays, only the globular regions of C1q inhibited the interaction between C1q and OmpK36 porin, demonstrating that C1q binds to porin through its globular region and not through the collagen-like stalks.
Albertí
S
S
Departamento de Biología Ambiental, Universidad de las Islas Baleares,Palma de Mallorca, Spain.
Marqués
G
G
Hernández-Allés
S
S
Rubires
X
X
Tomás
J M
JM
Vivanco
F
F
Benedí
V J
VJ
D001426:Bacterial Proteins; D001665:Binding Sites; D015922:Complement C1q / Q000276:immunology / Q000378:metabolism*; D003179:Complement C3b / Q000378:metabolism; D015938:Complement Membrane Attack Complex / Q000378:metabolism; D003171:Complement Pathway, Classical; D006801:Humans; D007526:Isoelectric Point; D007711:Klebsiella pneumoniae / Q000276:immunology*; D008969:Molecular Sequence Data; D018272:Porins / Q000276:immunology / Q000378:metabolism*
D001426:Bacterial Proteins; D015938:Complement Membrane Attack Complex; C092233:OmpK36 protein, Klebsiella pneumoniae; D018272:Porins; D015922:Complement C1q; D003179:Complement C3b