Interactions of the fatty acid-binding protein ReP1-NCXSQ with lipid membranes. Influence of the membrane electric field on binding and orientation

Galassi, Vanesa Viviana; Villarreal, Marcos Ariel; Posada, Velia; Montich, Guillermo G.

doi:10.1016/j.bbamem.2013.11.008

Cited by 10 publications

(21 citation statements)

References 45 publications

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“…These differences may influence the orientation of leishmanolysins along the membrane. Electrostatics indeed play a decisive role in protein orientation, as demonstrated for other proteins using experimental and theoretical methods ( Zamarreño et al 2012 , Galassi et al 2014 ). We hypothesize that the surface of Leishmania promastigotes is therefore composed of leishmanolysins adopting a broad range of orientations close to the membrane, therefore introducing an extra complexity that may favor the interactions with host cells.…”

Section: Discussionmentioning

confidence: 77%

Structural insights into leishmanolysins encoded on chromosome 10 of Leishmania (Viannia) braziliensis

Sutter¹,

Antunes²,

Silva-Almeida

et al. 2017

Mem. Inst. Oswaldo Cruz

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BACKGROUND Leishmanolysins have been described as important parasite virulence factors because of their roles in the infection of promastigotes and resistance to host’s defenses. Leishmania (Viannia) braziliensis contains several leishmanolysin genes in its genome, especially in chromosome 10. However, the functional impact of such diversity is not understood, but may be attributed partially to the lack of structural data for proteins from this parasite.OBJECTIVES This works aims to compare leishmanolysin sequences from L. (V.) braziliensis and to understand how the diversity impacts in their structural and dynamic features.METHODS Leishmanolysin sequences were retrieved from GeneDB. Subsequently, 3D models were built using comparative modeling methods and their dynamical behavior was studied using molecular dynamic simulations.FINDINGS We identified three subgroups of leishmanolysins according to sequence variations. These differences directly affect the electrostatic properties of leishmanolysins and the geometry of their active sites. We identified two levels of structural heterogeneity that might be related to the ability of promastigotes to interact with a broad range of substrates.MAIN CONCLUSION Altogether, the structural plasticity of leishmanolysins may constitute an important evolutionary adaptation rarely explored when considering the virulence of L. (V.) braziliensis parasites.

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Section: Discussionmentioning

confidence: 77%

Structural insights into leishmanolysins encoded on chromosome 10 of Leishmania (Viannia) braziliensis

Sutter¹,

Antunes²,

Silva-Almeida

et al. 2017

Mem. Inst. Oswaldo Cruz

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“…Fig 4A shows the trajectories of z L-P for ReP1-NCXSQ and L-BABP in the presence of EDMPC cationic membranes. For comparison, we also show in Fig 4B the same property computed for ReP1-NCXSQ in anionic membranes [ 8 ]. Both proteins migrated to the membrane and reached a constant distance within 1.75 and 2.75 nm.…”

Section: Resultsmentioning

confidence: 74%

“…Purified L-BABP was kindly supplied by Dr. Hugo Monaco and stored in 2 mM phosphate buffer, pH 7.5, at -70 o C. Recombinant ReP1-NCXSQ was expressed and purified as in Galassi et al [ 8 ].…”

Section: Methodsmentioning

confidence: 99%

“…We attempted to prove the relevance of electrostatic interactions in defining the orientation of peripheral membrane proteins. We have previously studied the binding to lipid membranes of the chicken liver bile acid-binding protein [ 6 ], (L-BABP, PDB ID 1TVQ [ 7 ]) and the regulatory protein of the squid nerve sodium calcium exchanger [ 8 ], (ReP1-NCXSQ, PDB 3PPT [ 9 ]). They share the common fold of the family: a β-barrel delimiting an inner cavity and a portal region with two α-helix segments.…”

Section: Introductionmentioning

confidence: 99%

“…They share the common fold of the family: a β-barrel delimiting an inner cavity and a portal region with two α-helix segments. We demonstrated, using binding assays and Molecular Dynamics (MD) simulations, that they interact with negatively charged lipid membranes and that electrostatic interactions have a major role on determining their orientation in the interface [ 6 , 8 , 10 – 13 ]. We have shown that the alignment of the dipole moment with the membrane electric field has a dominant role in the protein orientation within the interface.…”

Section: Introductionmentioning

confidence: 99%

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Relevance of the protein macrodipole in the membrane-binding process. Interactions of fatty-acid binding proteins with cationic lipid membranes

2018

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The fatty acid-binding proteins L-BABP and Rep1-NCXSQ bind to anionic lipid membranes by electrostatic interactions. According to Molecular Dynamics (MD) simulations, the interaction of the protein macrodipole with the membrane electric field is a driving force for protein binding and orientation in the interface. To further explore this hypothesis, we studied the interactions of these proteins with cationic lipid membranes. As in the case of anionic lipid membranes, we found that both proteins, carrying a negative as well as a positive net charge, were bound to the positively charged membrane. Their major axis, those connecting the bottom of the β-barrel with the α-helix portal domain, were rotated about 180 degrees as compared with their orientations in the anionic lipid membranes. Fourier transform infrared (FTIR) spectroscopy of the proteins showed that the positively charged membranes were also able to induce conformational changes with a reduction of the β-strand proportion and an increase in α-helix secondary structure. Fatty acid-binding proteins (FABPs) are involved in several cell processes, such as maintaining lipid homeostasis in cells. They transport hydrophobic molecules in aqueous medium and deliver them into lipid membranes. Therefore, the interfacial orientation and conformation, both shown herein to be electrostatically determined, have a strong correlation with the specific mechanism by which each particular FABP exerts its biological function.

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Conformational changes, from β-strand to α-helix, of the fatty acid-binding protein ReP1-NCXSQ in anionic lipid membranes: dependence with the vesicle curvature

2017

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We studied the conformational changes of the fatty acid-binding protein ReP1-NCXSQ in the interface of anionic lipid membranes. ReP1-NCXSQ is an acidic protein that regulates the activity of the Na/Ca exchanger in squid axon. The structure is a flattened barrel composed of two orthogonal β-sheets delimiting an inner cavity and a domain of two α-helix segments arranged as a hairpin. FTIR and CD spectroscopy showed that the interactions with several anionic lipids in the form of small unilamellar vesicles (SUVs) induced an increase in the proportion of helix secondary structure. Lower amount or no increase in α-helix was observed upon the interaction with anionic lipids in the form of large unilamellar vesicles (LUVs). The exception was 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) that was equally efficien to to induce the conformational change both in SUVs and in LUVs. In solution, the infrared spectra of ReP1-NCXSQ at temperatures above the unfolding displayed a band at 1617 cm characteristic of aggregated strands. This band was not observed when the protein interacted with DMPG, indicating inhibition of aggregation in the interface. Similarly to the observed in L-BABP, another member of the fatty acid binding proteins, a conformational change in ReP1-NCXSQ was coupled to the gel to liquid-crystalline lipid phase transition.

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Interactions of the fatty acid-binding protein ReP1-NCXSQ with lipid membranes. Influence of the membrane electric field on binding and orientation

Cited by 10 publications

References 45 publications

Structural insights into leishmanolysins encoded on chromosome 10 of Leishmania (Viannia) braziliensis

Structural insights into leishmanolysins encoded on chromosome 10 of Leishmania (Viannia) braziliensis

Relevance of the protein macrodipole in the membrane-binding process. Interactions of fatty-acid binding proteins with cationic lipid membranes

Conformational changes, from β-strand to α-helix, of the fatty acid-binding protein ReP1-NCXSQ in anionic lipid membranes: dependence with the vesicle curvature

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