Interaction of the Dengue Virus Fusion Peptide with Membranes Assessed by NMR: The Essential Role of the Envelope Protein Trp101 for Membrane Fusion
ABSTRACT Dengue virus (DV) infection depends on a step of membrane fusion, which occurs in the acidic environment of the endosome. This process is mediated by virus surface envelope glycoprotein, in which the loop between residues D98-G112 is considered to be crucial, acting as a fusion peptide. Here, we have characterized functionally and structurally the interaction between the DV fusion peptide and different model membranes by fluorescence and NMR. Its interaction was strongest in dodecylphosphocholine (DPC) micelles and anionic phosphatidylcholine/phosphatidylglycerol vesicles, the only vesicle that was fused by DV fusion peptide. The three-dimensional structure of DV fusion peptide bound to DPC micelles was solved by solution homonuclear NMR with an r.m.s.d. of 0.98 A. The most striking result obtained from the solution structure was the hydrophobic triad formed by residues W101, L107, and F108, pointing toward the same direction, keeping the segment between G102 and G106 in a loop conformation. The interaction of DV fusion peptide with phosphatidylcholine/phosphatidylglycerol vesicles was also mapped by transfer-nuclear Overhauser enhancement (NOE) experiments, in which the majority of the NOE cross-peaks were from the hydrophobic triad, corroborating the DPC-bound structure. Substitution of the residue W101 by an alanine residue completely abolished membrane binding and, thus, fusion by the peptide and its NOE cross-peaks. In conclusion, the 15-residue DV fusion peptide has intrinsic ability to promote membrane fusion, most likely due to the hydrophobic interaction among the residues W101, L107, and F108, which maintains its loop in the correct spatial conformation.
Interaction of the Dengue Virus Fusion Peptide with Membranes Assessed by NMR: The Essential Role of the Envelope Protein Trp101 for Membrane Fusion - ResearchGate.
Brookhaven Women in Science Talk at Brookhaven Lab on Friday, 10/16: 'Microscopes and Biomolecules in the Toy Box'
October 7, 2015
UPTON, NY — Ana-Carolina Zeri of Brazilian Biosciences National Laboratory will give a talk, "Microscopes and Biomolecules in the Toy Box," at the U.S. Department of Energy's Brookhaven National Laboratory on Friday, October 16, at 4:30 p.m. in the Physics Department seminar room. Sponsored by Brookhaven Science Associates and Brookhaven Women in Science, the talk is free and open to the public. All visitors to the Laboratory 16 and older must bring a photo I.D.
The Brazilian Biosciences National Laboratory, LNBio, is part of a complex of national laboratories, Centro Nacional de Pesquisa em Energy e Materiais (CNPEM) in Brazil. The complex includes the site of the first Synchrotron Light Source built in the Southern Hemisphere, the "Laboratorio Nacional de Luz Sincrotron," and future site of the next generation source, Sirius the CPEM, and is also home to Laboratorio Nacional de Ciencia e Tecnologia do Bioetanol (CTBE) and the Laboratorrio Nacional de Nanotechnologia (LNNano).
For her talk at Brookhaven, Zeri will discuss her activities as a scientist and an educator, and how her team uses nuclear magnetic resonance spectroscopy (NMR), to study biological samples with applications in cancer research, bacterial growth machinery, and pathogen defense mechanisms. The NMR techniques enable scientists to determine the shape (at an atomic level) of molecules, such as proteins, and investigate minute differences in the composition of biofluids, such as saliva, blood plasma and urine.
Winner of an Eisenhower Fellowship to enhance science, technology, engineering, and math (STEM) educational programs, Zeri will also discuss her personal experiences and challenges as a woman in the world of science. She will also present how she has combined her research with her passion for education and the social advancement of young women. She is also interested in the portrayal of girls and women in the media and how this impacts their career aspirations.
Ana Zeri completed her Ph.D in the United States and returned to Brazil where she currently is a principal investigator at Brazilian Biosciences National Laboratory. In addition to her research using nuclear magnetic resonance spectroscopy, Zeri also volunteers as a leader in educational outreach programs. Her future plans include studying the importance of science education in public policy.
Charge inversion by polyvalent ions is thought to be highly relevant for the number of biological processes, including action of drugs 19, gene delivery 20, DNA condensation7, and viral packing 21, 22. .. .. We demonstrate that at small voltages the gramicidin A channel in asymmetrically charged membranes displays current rectification which is close to that of a typical solid-state diode.
So flipped PS and those negative charges that Peregrine has the rights to target 100%... Happy Thanksgiving to all
The Effect of pH on the Electrical Capacitance of Phosphatidylcholine–Phosphatidylserine System in Bilayer Lipid Membrane Monika Naumowiczcorresponding author and Zbigniew Artur Figaszewski Institute of Chemistry, University of Bialystok, Al. J. Pilsudskiego 11/4, 15-443 Bialystok, Poland Laboratory of Electrochemical Power Sources Faculty of Chemistry, University of Warsaw, Pasteur St. 1, 02-093 Warsaw, Poland Monika Naumowicz, Phone: +48-85-6647487, Fax: +48-85-6647489, Email: lp.ude.bwu@nakinom. corresponding authorCorresponding author. .. .. .. Abstract
This paper reports measurements on the pH dependence of the electrical capacitance of lipid membranes formed by 1:1 phosphatidylcholine-phosphatidylserine mixtures. A theoretical model was developed to describe this dependence, in which the contributions of functional groups (as the active centers of adsorption of the hydrogen and hydroxide ions) to the overall membrane capacitance were assumed to be additive. The proposed model was verified experimentally using electrochemical impedance spectroscopy. The theoretical predictions agreed with the experimental results over the measured pH range. A minimum corresponding to the isoelectric point appeared in both the theoretical equation and the experimental data. Keywords: Electrochemical impedance spectroscopy pH, Acid–base equilibria, Bilayer membrane, Phosphatidylcholine, Phosphatidylserine
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Conclusions
In conclusion, noninvasive EIS method was used to characterize the capacitive properties of the PC–PS bilayer and to provide a quantitative description of the acid–base equilibria at the interface separating the electrolyte solution and bilayer. The electrical capacitance of the analyzed bilayer had a minimum value around pH 4.2. The value of the capacitance increased as the pH of the solution decreased or increased.
Therefore, pH appears to play an important role in setting the electrical properties of bilayer lipid membranes. This information may be useful for investigating the functional properties of lipid membranes and membrane proteins under aqueous pH stress.