Neisseria meningitidis Opc invasin binds to the cytoskeletal protein alpha-actinin.
Neisseria meningitidis Opc protein is an effective invasin for human endothelial cells. We have investigated novel human endothelial receptors targeted by Opc and observed that Opc-expressing bacteria interacted with a 100 kDa protein in whole-cell lysates of human endothelial and epithelial cells. The identity of the protein was established as alpha-actinin by mass spectrometry. Opc expression was essential for the recognition of alpha-actinin whether provided in a purified form or in cell extracts. The interaction of the two proteins did not involve intermediate molecules. As there was no demonstrable expression of alpha-actinin on the surfaces of any of the eight cell lines studied, the likelihood of the interactions after meningococcal internalization was examined. Confocal imaging demonstrated considerable colocalization of N. meningitidis with alpha-actinin especially after a prolonged period of internalization. This may imply that bacteria and alpha-actinin initially occur in separate compartments and co-compartmentalization occurs progressively over the 8 h infection period used. In conclusion, these studies have identified a novel and an intracellular target for the N. meningitidis Opc invasin. Since alpha-actinin is a modulator of a variety of signalling pathways and of cytoskeletal functions, its targeting by Opc may enable bacteria to survive/translocate across endothelial barriers.
Sa E Cunha
Claudia
C
Department of Cellular and Molecular Medicine, School of Medical Sciences, University of Bristol, Bristol, UK.
Griffiths
Natalie J
NJ
Murillo
Isabel
I
Virji
Mumtaz
M
D000185:Actinin / Q000378:metabolism*; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D042783:Endothelial Cells / Q000382:microbiology; D004847:Epithelial Cells; D006801:Humans; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D009345:Neisseria meningitidis / Q000502:physiology*; D011485:Protein Binding
D001425:Bacterial Outer Membrane Proteins; D003598:Cytoskeletal Proteins; D000185:Actinin; C074517:opc protein, bacteria