Document Detail


Time-resolved EPR immersion depth studies of a transmembrane peptide incorporated into bicelles.
MedLine Citation:
PMID:  22100865     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The reduction in EPR signal intensity of nitroxide spin-labels by ascorbic acid has been measured as a function of time to investigate the immersion depth of the spin-labeled M2δ AChR peptide incorporated into a bicelle system utilizing EPR spectroscopy. The corresponding decay curves of n-DSA (n=5, 7, 12, and 16) EPR signals have been used to (1) calibrate the depth of the bicelle membrane and (2) establish a calibration curve for measuring the depth of spin-labeled transmembrane peptides. The kinetic EPR data of CLS, n-DSA (n=5, 7, 12, and 16), and M2δ AChR peptide spin-labeled at Glu-1 and Ala-12 revealed excellent exponential and linear fits. For a model M2δ AChR peptide, the depth of immersion was calculated to be 5.8Å and 3Å for Glu-1, and 21.7Å and 19Å for Ala-12 in the gel-phase (298K) and L(α)-phases (318K), respectively. The immersion depth values are consistent with the pitch of an α-helix and the structural model of M2δ AChR incorporated into the bicelle system is in a good agreement with previous studies. Therefore, this EPR time-resolved kinetic technique provides a new reliable method to determine the immersion depth of membrane-bound peptides, as well as, explore the structural characteristics of the M2δ AChR peptide.
Authors:
Nisreen A Nusair; Daniel J Mayo; Tia D Dorozenski; Thomas B Cardon; Johnson J Inbaraj; Ethan S Karp; Justin P Newstadt; Stuart M Grosser; Gary A Lorigan
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2011-11-11
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  1818     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  2012 Mar 
Date Detail:
Created Date:  2012-02-06     Completed Date:  2012-04-24     Revised Date:  2014-09-17    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  821-8     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 Elsevier B.V. All rights reserved.
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MeSH Terms
Descriptor/Qualifier:
Animals
Electron Spin Resonance Spectroscopy / methods
Humans
Liposomes / chemistry*
Models, Molecular*
Peptides / chemistry*
Protein Structure, Secondary
Receptor, Muscarinic M2 / chemistry*
Spin Labels
Grant Support
ID/Acronym/Agency:
GM60259-01/GM/NIGMS NIH HHS; R01 GM080542/GM/NIGMS NIH HHS; R01 GM080542-01A2/GM/NIGMS NIH HHS; R01 GM080542-02S1/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Liposomes; 0/Peptides; 0/Receptor, Muscarinic M2; 0/Spin Labels
Comments/Corrections

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