Quantification of Protein-Lipid Selectivity using FRET: Application to the M13 Major Coat Protein

Biophysical Journal
Fábio FernandesM Prieto

Abstract

Quantification of lipid selectivity by membrane proteins has been previously addressed mainly from electron spin resonance studies. We present here a new methodology for quantification of protein-lipid selectivity based on fluorescence resonance energy transfer. A mutant of M13 major coat protein was labeled with 7-diethylamino-3((4'iodoacetyl)amino)phenyl-4-methylcoumarin to be used as the donor in energy transfer studies. Phospholipids labeled with N-(7-nitro-2-1,3-benzoxadiazol-4-yl) were selected as the acceptors. The dependence of protein-lipid selectivity on both hydrophobic mismatch and headgroup family was determined. M13 major coat protein exhibited larger selectivity toward phospholipids which allow for a better hydrophobic matching. Increased selectivity was also observed for anionic phospholipids and the relative association constants agreed with the ones already presented in the literature and obtained through electron spin resonance studies. This result led us to conclude that fluorescence resonance energy transfer is a promising methodology in protein-lipid selectivity studies.

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Citations

Aug 30, 2008·Chemistry and Physics of Lipids·Rodrigo F M de AlmeidaManuel Prieto
Jan 18, 2008·Biophysical Journal·Fábio FernandesManuel Prieto
Nov 19, 2008·Biochimica Et Biophysica Acta·Luís M S LouraManuel Prieto
Feb 22, 2005·Current Opinion in Biotechnology·Horst Wallrabe, Ammasi Periasamy
May 22, 2010·Biochimica Et Biophysica Acta·Laura PicasLuís M S Loura
May 16, 2014·Molecular Membrane Biology·Carme Suárez-GermàLuís M S Loura
Mar 15, 2015·Biochimica Et Biophysica Acta·Fábio FernandesLuís M S Loura
Jul 21, 2010·Biophysical Journal·Joel H NyströmThomas K M Nyholm
Aug 15, 2006·Biophysical Journal·David StoparMarcus A Hemminga
Mar 17, 2007·Biochimica Et Biophysica Acta·H RaghuramanAmitabha Chattopadhyay
Dec 12, 2007·Journal of Peptide Science : an Official Publication of the European Peptide Society·Renske W HesselinkManuel Prieto
Mar 20, 2010·European Biophysics Journal : EBJ·Luís M S LouraFábio Fernandes
Mar 30, 2017·Methods and Applications in Fluorescence·Valeriya M TrusovaNikolai Gadjev
May 3, 2021·Biochimica Et Biophysica Acta. Biomembranes·Mikito NakanoMichio Murata
May 8, 2013·The Journal of Physical Chemistry. B·Carme Suárez-GermàJordi Hernández-Borrell
Feb 3, 2012·The Journal of Physical Chemistry. B·Carme Suárez-GermàJordi Hernández-Borrell
Nov 10, 2012·The Journal of Physical Chemistry. B·Carme Suárez-GermàJordi Hernández-Borrell

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