Ionic strength-dependent alterations of membrane structure of red blood cells

1986 ◽  
Vol 6 (11) ◽  
pp. 1007-1015 ◽  
Author(s):  
Andreas Herrmann ◽  
Peter Müller
Keyword(s):  
Ionic Strength ◽  
Membrane Structure ◽  
Spin Labels ◽  
Head Group ◽  
Polar Head Group ◽  
Paramagnetic Resonance ◽  
Group A ◽  
Hydrophobic Tail ◽  
Low Ionic Strength ◽  
Electron Paramagnetic

Electron paramagnetic resonance (EPR) measurements using various fatty acid spin labels were performed on membranes of intact human erythrocytes at physiological, and at low ionic strength. In the case of spin probes bearing the nitroxide near the polar head group, a less restricted motion at low ionic strength was seen than with those labels with a nitroxide deeper within the hydrophobic tail of the membrane. Although these data clearly show an influence of ionic strength on membrane structure, and possibly a modified protein-lipid interaction, they cannot be simply discussed in terms of an altered membrane fluidity.

scholarly journals Studies of transmembrane peptides by pulse dipolar spectroscopy with semi-rigid TOPP spin labels

2021 ◽  
Author(s):  
Igor Tkach ◽  
Ulf Diederichsen ◽  
Marina Bennati
Keyword(s):  
Molecular Level ◽  
Dipolar Interaction ◽  
Spin Labels ◽  
Paramagnetic Centers ◽  
Structural Adaptation ◽  
Paramagnetic Resonance ◽  
Transmembrane Peptides ◽  
Distance Distributions ◽  
Lipid Environment ◽  
Electron Paramagnetic

AbstractElectron paramagnetic resonance (EPR)-based pulsed dipolar spectroscopy measures the dipolar interaction between paramagnetic centers that are separated by distances in the range of about 1.5–10 nm. Its application to transmembrane (TM) peptides in combination with modern spin labelling techniques provides a valuable tool to study peptide-to-lipid interactions at a molecular level, which permits access to key parameters characterizing the structural adaptation of model peptides incorporated in natural membranes. In this mini-review, we summarize our approach for distance and orientation measurements in lipid environment using novel semi-rigid TOPP [4-(3,3,5,5-tetramethyl-2,6-dioxo-4-oxylpiperazin-1-yl)-L-phenylglycine] labels specifically designed for incorporation in TM peptides. TOPP labels can report single peak distance distributions with sub-angstrom resolution, thus offering new capabilities for a variety of TM peptide investigations, such as monitoring of various helix conformations or measuring of tilt angles in membranes. Graphical Abstract

scholarly journals 2′-Alkynyl spin-labelling is a minimally perturbing tool for DNA structural analysis

2020 ◽  
Vol 48 (6) ◽  
pp. 2830-2840 ◽  
Author(s):  
Jack S Hardwick ◽  
Marius M Haugland ◽  
Afaf H El-Sagheer ◽  
Denis Ptchelkine ◽  
Frank R Beierlein ◽  
...  
Keyword(s):  
Epr Spectroscopy ◽  
Spin Labels ◽  
Resonance Spectroscopy ◽  
Conformational Ensemble ◽  
Dna Duplexes ◽  
Paramagnetic Resonance ◽  
Dynamics Simulations ◽  
Electron Paramagnetic ◽  
Spin Labelling

Abstract The determination of distances between specific points in nucleic acids is essential to understanding their behaviour at the molecular level. The ability to measure distances of 2–10 nm is particularly important: deformations arising from protein binding commonly fall within this range, but the reliable measurement of such distances for a conformational ensemble remains a significant challenge. Using several techniques, we show that electron paramagnetic resonance (EPR) spectroscopy of oligonucleotides spin-labelled with triazole-appended nitroxides at the 2′ position offers a robust and minimally perturbing tool for obtaining such measurements. For two nitroxides, we present results from EPR spectroscopy, X-ray crystal structures of B-form spin-labelled DNA duplexes, molecular dynamics simulations and nuclear magnetic resonance spectroscopy. These four methods are mutually supportive, and pinpoint the locations of the spin labels on the duplexes. In doing so, this work establishes 2′-alkynyl nitroxide spin-labelling as a minimally perturbing method for probing DNA conformation.

The inhibitory effects of polyoxyethylene detergents on human erythrocyte acetylcholinesterase and Ca2+ + Mg2+ ATPase

1989 ◽  
Vol 67 (2-3) ◽  
pp. 137-146 ◽  
Author(s):  
R. Blaine Moore ◽  
J. F. Manery ◽  
J. Still ◽  
V. N. Mankad
Keyword(s):  
Human Erythrocyte ◽  
Acetylcholinesterase Activity ◽  
Significant Inhibition ◽  
Erythrocyte Membranes ◽  
Head Group ◽  
Polar Head Group ◽  
Detergent Concentration ◽  
Human Erythrocyte Membranes ◽  
Hydrophobic Tail ◽  
Triton X

The activities of acetylcholinesterase and Ca2+ + Mg2+ ATPase were measured following treatment of human erythrocyte membranes with nonsolubilizing and solubilizing concentrations of Triton X-100. A concentration of 0.1% (v/v) Triton X-100 caused a significant inhibition of both enzymes. The inhibition appears to be caused by perturbations in the membrane induced by Triton X-100 incorporation. No acetylcholinesterase activity and little Ca2+ + Mg2+ ATPase activity were detected in the supernatant at 0.05% Triton X-100 although this same detergent concentration induced changes in the turbidity of the membrane suspension. Also, no inhibition of soluble acetylcholinesterase was observed over the entire detergent concentration range. The inhibition of these enzymes at 0.1% Triton X-100 was present over an eightfold range of membrane protein in the assay indicating an independence of the protein/detergent ratio. The losses in activities of these two enzymes could be prevented by either including phosphatidylserine in the Triton X-100 suspension or using Brij 96 which has the same polyoxyethylene polar head group but an oleyl hydrophobic tail instead of the p-tert-octylphenol group of Triton X-100. The results are discussed in regard to the differential recovery of enzyme activities over the entire detergent concentration range.Key words: Triton X-100, erythrocyte membranes, acetylcholinesterase, Ca2+ + Mg2+ ATPase, polyoxyethylene detergents.

scholarly journals Triarylmethyl Radicals: An EPR Study of 13C Hyperfine Coupling Constants

2017 ◽  
Vol 231 (4) ◽  
Author(s):  
Andrey A. Kuzhelev ◽  
Victor M. Tormyshev ◽  
Olga Yu. Rogozhnikova ◽  
Dmitry V. Trukhin ◽  
Tatiana I. Troitskaya ◽  
...  
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Epr Spectroscopy ◽  
Hyperfine Coupling ◽  
Coupling Constants ◽  
Spin Labels ◽  
Spin Probes ◽  
Paramagnetic Resonance ◽  
Hyperfine Coupling Constants ◽  
Electron Paramagnetic ◽  
Triarylmethyl Radicals

AbstractTriarylmethyl (TAM) radicals are widely used in electron paramagnetic resonance (EPR) spectroscopy as spin labels and in EPR imaging as spin probes for

scholarly journals DNA complexes with human apurinic/apyrimidinic endonuclease 1: structural insights revealed by pulsed dipolar EPR with orthogonal spin labeling

2019 ◽  
Vol 47 (15) ◽  
pp. 7767-7780 ◽  
Author(s):  
Olesya A Krumkacheva ◽  
Georgiy Yu Shevelev ◽  
Alexander A Lomzov ◽  
Nadezhda S Dyrkheeva ◽  
Andrey A Kuzhelev ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Md Simulations ◽  
Dna Lesions ◽  
Spin Labels ◽  
Paramagnetic Resonance ◽  
Applied Pulse ◽  
Oligonucleotide Duplex ◽  
Nitroxide Spin Labels ◽  
Ap Sites ◽  
Electron Paramagnetic

Abstract A DNA molecule is under continuous influence of endogenous and exogenous damaging factors, which produce a variety of DNA lesions. Apurinic/apyrimidinic sites (abasic or AP sites) are among the most common DNA lesions. In this work, we applied pulse dipolar electron paramagnetic resonance (EPR) spectroscopy in combination with molecular dynamics (MD) simulations to investigate in-depth conformational changes in DNA containing an AP site and in a complex of this DNA with AP endonuclease 1 (APE1). For this purpose, triarylmethyl (TAM)-based spin labels were attached to the 5′ ends of an oligonucleotide duplex, and nitroxide spin labels were introduced into APE1. In this way, we created a system that enabled monitoring the conformational changes of the main APE1 substrate by EPR. In addition, we were able to trace substrate-to-product transformation in this system. The use of different (orthogonal) spin labels in the enzyme and in the DNA substrate has a crucial advantage allowing for detailed investigation of local damage and conformational changes in AP-DNA alone and in its complex with APE1.

Nitroxides and nucleic acids: Chemistry and electron paramagnetic resonance (EPR) spectroscopy

2011 ◽  
Vol 83 (3) ◽  
pp. 677-686 ◽  
Author(s):  
Snorri Th. Sigurdsson
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Nucleic Acids ◽  
Epr Spectroscopy ◽  
Noncovalent Interactions ◽  
Spin Labeling ◽  
Structural Features ◽  
Spin Labels ◽  
Abasic Site ◽  
Paramagnetic Resonance ◽  
Electron Paramagnetic

Electron paramagnetic resonance (EPR) spectroscopy has increasingly been applied for the study of nucleic acid structure and dynamics. Such studies require incorporation of free radicals (spin labels) into the biopolymer. The labels can be incorporated during chemical synthesis of the oligomer (phosphoramidite approach) or postsynthetically, by reaction of a spin-labeling reagent with a reactive functional group on the oligonucleotide. Incorporation of the rigid nitroxide spin label Ç is an example of the phosphoramidite method, and reaction of a spin-labeled azide with an alkyne-modified oligomer to yield a triazole-derived, spin-labeled nucleotide illustrates the postsynthetic spin-labeling strategy. Characterization and application of these labels to study structural features of DNA by EPR spectroscopy is discussed. Finally, a new spin-labeling strategy is described for nucleic acids that relies on noncovalent interactions between a spin-labeled nucleobase and an abasic site in duplex DNA.

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