Site-Directed Spin-Labeling of Nucleic Acids by Click Chemistry: Detection of Abasic Sites in Duplex DNA by EPR Spectroscopy

2010 ◽  
Vol 132 (30) ◽  
pp. 10424-10428 ◽  
Author(s):  
Ulla Jakobsen ◽  
Sandip A. Shelke ◽  
Stefan Vogel ◽  
Snorri Th. Sigurdsson
Keyword(s):  
Nucleic Acids ◽  
Click Chemistry ◽  
Epr Spectroscopy ◽  
Spin Labeling ◽  
Duplex Dna ◽  
Abasic Sites ◽  
Site Directed Spin Labeling

Site-directed spin labeling EPR spectroscopy in protein research

2013 ◽  
Vol 394 (10) ◽  
pp. 1281-1300 ◽  
Author(s):  
Johann P. Klare
Keyword(s):  
Nucleic Acids ◽  
Epr Spectroscopy ◽  
Protein Function ◽  
Conformational Dynamics ◽  
Spin Labeling ◽  
Paramagnetic Resonance ◽  
Protein Research ◽  
Site Directed Spin Labeling ◽  
Electron Paramagnetic ◽  
Natronomonas Pharaonis

Abstract Site-directed spin labeling (SDSL) in combination with electron paramagnetic resonance (EPR) spectroscopy has emerged as an efficient tool to elucidate the structure and the conformational dynamics of proteins under conditions close to the native state. This review article summarizes the basics as well as the recent progress in SDSL and EPR methods, especially for investigations on protein structure, protein function, and interaction of proteins with other proteins or nucleic acids. Labeling techniques as well as EPR methods are introduced and exemplified with applications to systems that have been studied in the author’s laboratory in the past 15 years, headmost the sensory rhodopsin-transducer complex mediating the photophobic response of the halophilic archaeum Natronomonas pharaonis. Further examples underline the application of SDSL EPR spectroscopy to answer specific questions about the system under investigation, such as the nature and influence of interactions of proteins with other proteins or nucleic acids. Finally, it is discussed how SDSL EPR can be combined with other biophysical techniques to combine the strengths of the different methodologies.

scholarly journals Posttranscriptional site-directed spin labeling of large RNAs with an unnatural base pair system under non-denaturing conditions

2020 ◽  
Vol 11 (35) ◽  
pp. 9655-9664
Author(s):  
Yan Wang ◽  
Venkatesan Kathiresan ◽  
Yaoyi Chen ◽  
Yanping Hu ◽  
Wei Jiang ◽  
...  
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Epr Spectroscopy ◽  
Base Pair ◽  
Spin Labeling ◽  
Paramagnetic Resonance ◽  
Site Directed Spin Labeling ◽  
Electron Paramagnetic

Site-directed spin labeling (SDSL) of large RNAs for electron paramagnetic resonance (EPR) spectroscopy has remained challenging to date.

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.

Estimation of binding parameters for the protein–protein interaction using a site-directed spin labeling and EPR spectroscopy

2007 ◽  
Vol 37 (4) ◽  
pp. 483-493 ◽  
Author(s):  
Marcin Sarewicz ◽  
Sebastian Szytuła ◽  
Małgorzata Dutka ◽  
Artur Osyczka ◽  
Wojciech Froncisz
Keyword(s):  
Protein Interaction ◽  
Epr Spectroscopy ◽  
Spin Labeling ◽  
Binding Parameters ◽  
Protein Protein Interaction ◽  
Site Directed Spin Labeling ◽  
A Site

scholarly journals Site-directed spin labeling measurements of nanometer distances in nucleic acids using a sequence-independent nitroxide probe

2006 ◽  
Vol 34 (17) ◽  
pp. 4722-4730 ◽  
Author(s):  
Qi Cai ◽  
Ana Karin Kusnetzow ◽  
Wayne L. Hubbell ◽  
Ian S. Haworth ◽  
Gian Paola C. Gacho ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Spin Labeling ◽  
Nitroxide Probe ◽  
Site Directed Spin Labeling
Sign in / Sign up

Export Citation Format

Share Document