Reference : Nanobodies as structural probes to investigate the mechanism of fibril formation by the ...
Scientific congresses and symposiums : Poster
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/104040
Nanobodies as structural probes to investigate the mechanism of fibril formation by the amyloidogenic variants of human lysozyme.
English
Dumont, Janice mailto [Université de Liège - ULg > Département des sciences de la vie > Enzymologie et repliement des protéines >]
Menzer, Linda [Université de Liège - ULg > > > 2e an. master bioch. & biol. moléc. & cell., fin. appr.]
Pardon, Els [Vrije Universiteit Brussel - VUB > Departement ultrastructure > > >]
Wyns, Lode [Vrije Universiteit Brussel - VUB > > > >]
Steyaert, Jan [Vrije Universiteit Brussel - VUB > Departement Ultrastructure > > >]
Dobson, Christopher [University of Cambridge > Departement of chemistry > > >]
Dumoulin, Mireille mailto [Université de Liège - ULg > Département des sciences de la vie > Enzymologie et repliement des protéines >]
2010
No
International
Single Domain Antibodies Come of Age
14-15 octobre 2010
Ablynx
Ghent
Belgique
[en] Amyloid Fibril ; Nanobodies ; Lysozyme
[en] Six variants of human lysozyme (single-point mutations I56T, F57I, W64R, D67H and double
mutations F57I/T70N, W112R/T70N) are associated with a hereditary non-neuropathic systemic
amyloidose. These proteins form extracellular amyloid fibrils that deposit in a wide range of tissues
and organs such as liver, spleen and kidneys where they cause damages [1]. It was shown that the D67H and I56T mutations cause a loss in stability and more particularly a loss of global cooperativity of protein [1]. Consequently, under physiologically relevant conditions, these variants can transiently populate a partially unfolded state in which the beta-domain and the C-helix are cooperatively unfolded while the rest of the protein remains native like [1]. The formation of intermolecular interactions between the regions that are unfolded in this intermediate state is likely to be a fundamental trigger of the aggregation process that ultimately leads to the formation and deposition of fibrils in tissues. The binding of three variable domain of camelid antibodies – also named nanobodies - (cAb-HuL 6 [2], cAb-HuL 5 and cAb-HuL 22 [3]) raised against the wild type human lysozyme inhibit in vitro the formation of amyloid fibrils by the lysozyme variants. These three nanobodies bind on different regions of lysozyme and act as Amyloid fibrils inhibitor through different mechanisms. On one hand, cAb-HuL 6 and cAb-HuL 22 stabilize the native state of the lysozyme variants thus restoring the global cooperativity characteristic of the wild-type protein. On the other, cAb-HuL 5 probably acts by binding soluble prefibrillar aggregates. In the present work, sixteen other nanobodies specific of human lysozyme have been generated. Competition experiments have shown that they bind to five non overlapping epitopes. The effects of the binding of these nanobodies on the stability of the D67H variant of human lysozyme and on its aggregation into amyloid fibrils will be discussed.
http://hdl.handle.net/2268/104040

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