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

  • Vanesa V. Galassi
  • Silvina R. Salinas
  • Guillermo G. Montich
Original Article

Abstract

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+/Ca2+ 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−1 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.

Keywords

ReP1-NCXSQ Lipid membrane Protein conformational change Infrared spectroscopy Circular dichroism Membrane curvature 

Abbreviations

ReP1-NCXSQ

Regulatory protein of the squid nerve sodium calcium exchanger

FABP

Fatty acid-binding protein

L-BABP

Chicken liver bile acid-binding protein

DMPG

1,2-dimyristoyl-sn-glycero-3-phosphoglycerol

DPPG

1,2-dipalmytoyl-sn-glycero-3-phosphoglycerol

POPG

1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol

DMPA

1,2-dimyristoyl-sn-glycero-3-phosphate

DMPS

1,2-dimyristoyl-sn-glycero-3-phosphoserine

DPPA

1,2-dipalmitoyl -sn-glycero-3-phosphate

DLPA

1,2-dilauroyl-sn-glycero-3-phosphate

DMPC

1,2-dimyristoyl-sn-glycero-3-phosphocholine

LUV

Large unilamellar vesicle

FTIR

Fourier transform infrared

CD

Circular dichroism

FSD

Fourier self-deconvolution

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Copyright information

© European Biophysical Societies' Association 2017

Authors and Affiliations

  • Vanesa V. Galassi
    • 1
    • 2
    • 3
  • Silvina R. Salinas
    • 1
    • 2
    • 4
  • Guillermo G. Montich
    • 1
    • 2
  1. 1.Departamento de Química Biológica “Ranwel Caputto”, Facultad de Ciencias QuímicasUniversidad Nacional de CórdobaCórdobaArgentina
  2. 2.Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC)CONICET, Universidad Nacional de CórdobaCórdobaArgentina
  3. 3.CONICET, Facultad de Ciencias Exactas y NaturalesUniversidad Nacional de CuyoMendoza Argentina
  4. 4.Centro de Excelencia en Productos y Procesos Córdoba-CONICET, Pabellón CEPROCOR, X5164CórdobaArgentina

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