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Azurin interaction with the lipid raft components ganglioside GM-1 and caveolin-1 increases membrane fluidity and sensitivity to anti-cancer drugs

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Version 2 2018-08-05, 06:51
Version 1 2018-07-02, 12:46
journal contribution
posted on 2018-08-05, 06:51 authored by Nuno Bernardes, Ana Rita Garizo, Sandra N. Pinto, Bernardo Caniço, Catarina Perdigão, Fábio Fernandes, Arsenio M. Fialho

Membrane lipid rafts are highly ordered microdomains and essential components of plasma membranes. In this work, we demonstrate that azurin uptake by cancer cells is, in part, mediated by caveolin-1 and GM-1, lipid rafts’ markers. This recognition is mediated by a surface exposed hydrophobic core displayed by azurin since the substitution of a phenylalanine residue in position 114 facing the hydrophobic cavity by alanine impacts such interactions, debilitating the uptake of azurin by cancer cells. Treating of cancer cells with azurin leads to a sequence of events: alters the lipid raft exposure at plasma membranes, causes a decrease in the plasma membrane order as examined by Laurdan two-photon imaging and leads to a decrease in the levels of caveolin-1. Caveolae, a subset of lipid rafts characterized by the presence of caveolin-1, are gaining increasing recognition as mediators in tumor progression and resistance to standard therapies. We show that azurin inhibits growth of cancer cells expressing caveolin-1, and this inhibition is only partially observed with mutant azurin. Finally, the simultaneous administration of azurin with anticancer therapeutic drugs (paclitaxel and doxorubicin) results in an enhancement in their activity, contrary to the mutated protein.

Funding

This work was supported by the Fundação para a Ciência e a Tecnologia [PTDC/EBBBIO/100326/2008]; [FAPESP/20107/2014]; [UID/BIO/04565/2013]; [PPBI-POCI-01-0145-FEDER-022122] and from Programa Operacional Regional de Lisboa 2020 (Project N. 007317). Scholarships from Fundação para a Ciência e a Tecnologia [SFRH/BPD/98162/2013] to NB;;[SFRH/BD/122636/2016] to ARG; ; [SFRH/BPD/92409/2013] to SNP;and [IF/00386/2015]to FF are acknowledged.

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