Document Detail


Site-specific incorporation of nitroxide spin-labels into 2'-positions of nucleic acids.
MedLine Citation:
PMID:  17703207     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A protocol is described for the incorporation of nitroxide spin-labels into specific 2'-sites within nucleic acids. This labeling strategy facilitates the investigation of nucleic acid structure and dynamics using electron paramagnetic resonance (EPR) spectroscopy and macromolecular complex formation using paramagnetic relaxation enhancement NMR spectroscopy. A spin-labeling reagent, 4-isocyanato TEMPO, which can be prepared in one facile step or obtained commercially, is used for postsynthetic modification of site-specifically 2'-amino-modified nucleic acids. This spin-labeling protocol has been applied primarily to RNA, but is also applicable to DNA. Subsequently, EPR spectroscopic analysis of the spin-labeled nucleic acids allows for the measurements of distances, solvent accessibilities and conformation dynamics. Using the spin-labeling strategy described here, spin-labeled samples can be prepared in 2-4 d.
Authors:
Thomas E Edwards; Snorri Th Sigurdsson
Related Documents :
50567 - Possible association of oncornavirus type d with some forms of human cancer.
3793177 - Metabolic activation of vinyl chloride, formation of nucleic acid adducts and relevance...
12692537 - Enantiomeric interactions between nucleic acid bases and amino acids on solid surfaces.
11605637 - Survivability of biomolecules during extraterrestrial delivery: new results on pyrolysi...
16640447 - Similarity of salt influences on the ph of buffers, polyelectrolytes, and proteins.
8057847 - Evolution of antibiotic resistance: several different amino acid substitutions in an ac...
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nature protocols     Volume:  2     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2007  
Date Detail:
Created Date:  2007-08-17     Completed Date:  2007-12-06     Revised Date:  2008-03-24    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  1954-62     Citation Subset:  IM    
Affiliation:
University of Iceland, Science Institute, Dunhaga 3, Reykjavik, Iceland.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Cyclic N-Oxides
Electron Spin Resonance Spectroscopy / methods*
Electrophoresis, Polyacrylamide Gel
Nuclear Magnetic Resonance, Biomolecular
RNA, Catalytic / chemistry*
Spin Labels*
Chemical
Reg. No./Substance:
0/Cyclic N-Oxides; 0/RNA, Catalytic; 0/Spin Labels; 0/hammerhead ribozyme; 2564-83-2/TEMPO

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  A protocol for isolation and visualization of yeast nuclei by scanning electron microscopy (SEM).
Next Document:  Derivation of human embryonic stem cells from single blastomeres.