Fernández…Giraldo {Sci Rep 6: 23144}
The bacterial prionoid RepA-WH1 builds pores at model lipidic membranes. Left: mCherry-tagged RepA-WH1 (red) binds to and leaks the contents (calcein, green) of liposomes. Right: Average EM 2D-projection of a ring oligomer with its constituent RepA-WH1 monomers fitted.
RepA-WH1, the agent of an amyloid proteinopathy in bacteria, builds oligomeric pores through lipid vesicles
Fernández C, Núñez-Ramírez R, Jiménez M, Rivas G, Giraldo R.
Sci Rep 2016 Mar.; 6: 23144.
RepA-WH1 is a disease-unrelated protein that recapitulates in bacteria key aspects of human amyloid proteinopathies: i) It undergoes ligand-promoted amyloidogenesis in vitro; ii) its aggregates are able to seed/template amyloidosis on soluble protein molecules; iii) its conformation is modulated by Hsp70 chaperones in vivo, generating transmissible amyloid strains; and iv) causes proliferative senescence. Membrane disruption by amyloidogenic oligomers has been found for most proteins causing human neurodegenerative diseases. Here we report that, as for PrP prion and α-synuclein, acidic phospholipids also promote RepA-WH1 amyloidogenesis in vitro. RepA-WH1 molecules bind to liposomes, where the protein assembles oligomeric membrane pores. Fluorescent tracer molecules entrapped in the lumen of the vesicles leak through these pores and RepA-WH1 can then form large aggregates on the surface of the vesicles without inducing their lysis. These findings prove that it is feasible to generate in vitro a synthetic proteinopathy with a minimal set of cytomimetic components and support the view that cell membranes are primary targets in protein amyloidoses.
PubMed: 26984374. Doi: 10.1038/srep23144.
