Monica F Vidal

All Publications

• Membrane Partitioning: "Classical" and "Nonclassical" Hydrophobic Effects JOURNAL OF MEMBRANE BIOLOGY Fernandez-Vidal, M., White, S. H., Ladokhin, A. S. 2011; 239 (1-2): 5-14

  Abstract

  The free energy of transfer of nonpolar solutes from water to lipid bilayers is often dominated by a large negative enthalpy rather than the large positive entropy expected from the hydrophobic effect. This common observation has led to the idea that membrane partitioning is driven by the "nonclassical" hydrophobic effect. We examined this phenomenon by characterizing the partitioning of the well-studied peptide melittin using isothermal titration calorimetry (ITC) and circular dichroism (CD). We studied the temperature dependence of the entropic (-TΔS) and enthalpic (ΔH) components of free energy (ΔG) of partitioning of melittin into lipid membranes made of various mixtures of zwitterionic and anionic lipids. We found significant variations of the entropic and enthalpic components with temperature, lipid composition and vesicle size but only small changes in ΔG (entropy-enthalpy compensation). The heat capacity associated with partitioning had a large negative value of about -0.5 kcal mol(-1) K(-1). This hallmark of the hydrophobic effect was found to be independent of lipid composition. The measured heat capacity values were used to calculate the hydrophobic-effect free energy ΔG (hΦ), which we found to dominate melittin partitioning regardless of lipid composition. In the case of anionic membranes, additional free energy comes from coulombic attraction, which is characterized by a small effective peptide charge due to the lack of additivity of hydrophobic and electrostatic interactions in membrane interfaces [Ladokhin and White J Mol Biol 309:543-552, 2001]. Our results suggest that there is no need for a special effect-the nonclassical hydrophobic effect-to describe partitioning into lipid bilayers.

  View details for DOI 10.1007/s00232-010-9321-y

  View details for Web of Science ID 000286665000002

  View details for PubMedID 21140141

• CD Spectroscopy of Peptides and Proteins Bound to Large Unilamellar Vesicles JOURNAL OF MEMBRANE BIOLOGY Ladokhin, A. S., Fernandez-Vidal, M., White, S. H. 2010; 236 (3): 247-253

  Abstract

  Circular dichroism (CD) spectroscopy is an essential tool for determining the conformation of proteins and peptides in membranes. It can be particularly useful for measuring the free energy of partitioning of peptides into lipid vesicles. The belief is broadly held that such CD measurements can only be made using sonicated small unilamellar vesicles (SUVs) because light scattering associated with extruded large unilamellar vesicles (LUVs) is unacceptably high. We have examined this issue using several experimental approaches in which a chiral object (i.e., peptide or protein) is placed both on the membrane and outside the membrane. We show that accurate CD spectra can be collected in the presence of LUVs. This is important because SUVs, unlike LUVs, are metastable and consequently unsuitable for equilibrium thermodynamic measurements. Our data reveal that undistorted CD spectra of peptides can be measured at wavelengths above 200 nm in the presence of up to 3 mM LUVs and above 215 nm in the presence of up to 7 mM LUVs. We introduce a simple way of characterizing the effect on CD spectra of light scattering and absorption arising from suspensions of vesicles of any diameter. Using melittin as an example, we show that CD spectroscopy can be used to determine the fractional helical content of peptides in LUVs and to measure their free energy of partitioning of into LUVs.

  View details for DOI 10.1007/s00232-010-9291-0

  View details for Web of Science ID 000281794000002

  View details for PubMedID 20706833

• Insertion of short transmembrane helices by the Sec61 translocon PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Jaud, S., Fernandez-Vidal, M., Nilsson, I., Meindi-Beinker, N. M., Huebner, N. C., Tobias, D. J., von Heijne, G., White, S. H. 2009; 106 (28): 11588-11593

  Abstract

  The insertion efficiency of transmembrane (TM) helices by the Sec61 translocon depends on helix amino acid composition, the positions of the amino acids within the helix, and helix length. We have used an in vitro expression system to examine systematically the insertion efficiency of short polyleucine segments (L(n), n = 4 ... 12) flanked at either end by 4-residue sequences of the form XXPX-L(n)-XPXX with X = G, N, D, or K. Except for X = K, insertion efficiency (p) is <10% for n < 8, but rises steeply to 100% for n = 12. For X = K, p is already close to 100% for n = 10. A similar pattern is observed for synthetic peptides incorporated into oriented phospholipid bilayer arrays, consistent with the idea that recognition of TM segments by the translocon critically involves physical partitioning of nascent peptide chains into the lipid bilayer. Molecular dynamics simulations suggest that insertion efficiency is determined primarily by the energetic cost of distorting the bilayer in the vicinity of the TM helix. Very short lysine-flanked leucine segments can reduce the energetic cost by extensive hydrogen bonding with water and lipid phosphate groups (snorkeling) and by partial unfolding.

  View details for DOI 10.1073/pnas.0900638106

  View details for Web of Science ID 000267972700031

  View details for PubMedID 19581593

• Liposome destabilization induced by synthetic lipopeptides corresponding to envelope and non-structural domains of GBV-C/HGV virus. Conformational requirements for leakage BIOPHYSICAL CHEMISTRY Fernandez-Vidal, M., Rojo, N., Herrera, E., Gomara, M. J., Haro, I. 2008; 132 (1): 55-63

  Abstract

  Liposomes have been used primarily as a model system for studying biological membranes. Numerous chemical, biochemical and biophysical methods have been used to elucidate the various aspects of the interaction between proteins or peptides and phospholipids. Having in mind the potential use of synthetic lipopeptides as antiviral therapies and aiming for a better understanding of the molecular interaction of the GBV-C/HGV with liposomes as model membranes, epitopes of GBV-C/HGV located at the E2 (99-118) and NS3(440-460) regions were selected. Peptides were modified at the N-terminus with acyl chains of different length (C(14) and C(16)) yielding the corresponding myristoil and palmytoil lipopeptides. The main aim of the present study was to get insight into the membrane-interacting properties of the above-described synthetic lipopeptides and to study their inhibition of the capacity of perturbing model membranes of fusion peptide of HIV-1 using fluorescence spectroscopy. In an attempt to establish a relationship between peptide membrane activity and structure, we use Circular Dichroism (CD) and Fourier-Transform Infrared Spectroscopy (FTIR).

  View details for DOI 10.1016/j.bpe.2007.10.009

  View details for Web of Science ID 000251496700008

  View details for PubMedID 17988786

• 3D-Structure of the interior fusion peptide of HGV/GBV-C by 1H NMR, CD and molecular dynamics studies ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS Mazzini, S., Fernandez-Vidal, M., Galbusera, V., Castro-Roman, F., Bellucci, M. C., Ragg, E., Haro, I. 2007; 465 (1): 187-196

  Abstract

  In this work, we present a structural characterization of the putative fusion peptide E2(279-298) corresponding to the E2 envelope protein of the HGV/GBV-C virus by (1)H NMR, CD and MD studies performed in H(2)O/TFE and in lipid model membranes. The peptide is largely unstructured in water, whereas in H(2)O/TFE and in model membranes it adopts an helical structure (approximately 65-70%). The partitioning free energy DeltaG ranges from -6 to -7.5 kcal mol(-1). OCD measurements on peptide-containing hydrated and oriented lipid multilayers showed that the peptide adopts a predominantly surface orientation. The (1)H NMR data (observed NOEs, deuterium exchange rates, Halpha chemical shift index and vicinal coupling constants) and the molecular dynamics calculations support the conclusions that the peptide adopts a stable helix in the C-terminal 9-18 residues slightly inserted into the lipid bilayer and a major mobility in the amino terminus of the sequence (1-8 residues).

  View details for DOI 10.1016/j.abb.2007.05.024

  View details for Web of Science ID 000249176400022

  View details for PubMedID 17603997

• Folding amphipathic helices into membranes: Amphiphilicity trumps hydrophobicity JOURNAL OF MOLECULAR BIOLOGY Fernandez-Vidall, M., Jayasinghe, S., Ladokhin, A. S., White, S. H. 2007; 370 (3): 459-470

  Abstract

  High amphiphilicity is a hallmark of interfacial helices in membrane proteins and membrane-active peptides, such as toxins and antimicrobial peptides. Although there is general agreement that amphiphilicity is important for membrane-interface binding, an unanswered question is its importance relative to simple hydrophobicity-driven partitioning. We have examined this fundamental question using measurements of the interfacial partitioning of a family of 17-residue amidated-acetylated peptides into both neutral and anionic lipid vesicles. Composed only of Ala, Leu, and Gln residues, the amino acid sequences of the peptides were varied to change peptide amphiphilicity without changing total hydrophobicity. We found that peptide helicity in water and interface increased linearly with hydrophobic moment, as did the favorable peptide partitioning free energy. This observation provides simple tools for designing amphipathic helical peptides. Finally, our results show that helical amphiphilicity is far more important for interfacial binding than simple hydrophobicity.

  View details for DOI 10.1016/j.jmb.2007.05.016

  View details for Web of Science ID 000247904300006

  View details for PubMedID 17532340

• Application of a chimeric synthetic peptide in the development of a serologic method for the diagnosis of hepatitis G virus infection PROTEIN AND PEPTIDE LETTERS Fernandez-Vidal, M., CUBERO, M. D., Ercilla, G., Gomara, M. J., Haro, I. 2007; 14 (9): 865-870

  Abstract

  New putative antigenic peptides corresponding to the N- and C-terminal of the E2 envelope protein of GBV-C/HGV were synthesized using solid-phase chemistry. The antigens were obtained in linear and chimeric forms with the main aim of improving the sensitivity of the enzyme immunoassays. Furthermore, CD and FTIR have been used in conjunction to characterize their conformational changes showing that the chimeric peptide presents a more ordered secondary structure than its parent peptides.

  View details for Web of Science ID 000253577500006

  View details for PubMedID 18045227

• Structural, dynamic and mechanical properties of POPC at low cholesterol concentration studied in pressure/temperature space EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS Rappolt, M., Vidal, M. F., KRIECHBAUM, M., Steinhart, M., Amenitsch, H., Bernstorff, S., Laggner, P. 2003; 31 (8): 575-585

  Abstract

  We have studied the structural, dynamic and mechanical properties of 1-palmitoyl-2-oleoyl- sn-glycero-3-phosphatidylcholine (POPC)/cholesterol binary mixtures by small-angle X-ray scattering. Our investigations were concentrated on the biologically most relevant pressure-temperature-cholesterol regime, i.e. the liquid crystalline phase and its phase boundary to the lamellar gel phase within a cholesterol concentration up to 25 mol%. From the dependence of the transition pressure we derived a value of 19 kJ/mol for the transition enthalpy Delta H(m) of POPC in excess water. With increasing cholesterol concentration, Delta H(m) drops to about 7 kJ/mol at 20 mol% cholesterol. Time-resolved pressure-scan (p-scan) and temperature-jump (T-jump) experiments reveal that at low cholesterol content (<5-8 mol%) the fluidity and also the bilayer compressibility increase remarkably. In contrast, at concentrations between 5 and 25 mol% cholesterol the bilayer becomes again more rigid and the lipid bilayer spacing increases about 2 A. Theses changes are attributed to the onset of phase separation between liquid disordered and liquid ordered phases. The fluid-fluid miscibility gap for this mono-unsaturated lecithin species is strongly enlarged compared with saturated lecithins.

  View details for DOI 10.1007/s00249-002-0253-z

  View details for Web of Science ID 000181582100002

  View details for PubMedID 12582817