Document Detail


Two-dimensional electrophoresis with cationic detergents, a powerful tool for the proteomic analysis of myelin proteins. Part 1: technical aspects of electrophoresis.
MedLine Citation:
PMID:  17960830     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The analysis of proteins in damaged myelin is crucial to clarify the mechanisms of dysmyelination and demyelination. In the present study, proteomic analysis of myelin using a modified two-dimensional electrophoresis (2-DE) method was carried out to obtain a better understanding of myelin biology. Although standard 2-DE (immobilized pH gradient isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis; IPG/SDS-PAGE) methods of analysis provide high resolutions of soluble proteins with isoelectric focusing points in the pH range of 4-8, major myelin components include highly basic proteins are compacted at the basic edge of the 2-DE gels and are not sufficiently separated for satisfactory analysis. In an attempt to improve the separation of these proteins, an alternative 2-DE method using the cationic detergents was applied. In part 1 of this study, we describe technical aspects of conditioning 2-DE using cationic detergent. In the accompanying paper (part 2), practical 2-DE analysis using cationic detergents is described to identify proteins in the purified CNS myelin fraction. We carried out benzyldimethyl-n-hexadecylammonium chloride (16-BAC)/SDS-PAGE 2-DE and tested 2-DE with four other cationic detergents. We found that 16-BAC was the most effective agent for separation of myelin proteins and that hexadecyltrimethylammonium bromide (cetyltrimethylammonium bromide; CTAB) was the most effective agent for solubilization of myelin proteins. The combination of 16-BAC/SDS-PAGE and CTAB/SDS-PAGE is a powerful tool for the analysis of myelin proteins, including highly basic, high-MW (MW > 100K), and integral membrane proteins.
Authors:
Yoshihide Yamaguchi; Yudai Miyagi; Hiroko Baba
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of neuroscience research     Volume:  86     ISSN:  1097-4547     ISO Abbreviation:  J. Neurosci. Res.     Publication Date:  2008 Mar 
Date Detail:
Created Date:  2008-02-18     Completed Date:  2008-07-08     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7600111     Medline TA:  J Neurosci Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  755-65     Citation Subset:  IM    
Copyright Information:
(c) 2007 Wiley-Liss, Inc.
Affiliation:
Department of Molecular Neurobiology, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo, Japan. yoshiy@ps.toyaku.ac.jp
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Western
Brain Chemistry
Cations*
Detergents*
Electrophoresis, Gel, Two-Dimensional / methods*
Myelin Proteins / chemistry*,  isolation & purification
Myelin Sheath / chemistry
Proteomics / methods*
Rats
Chemical
Reg. No./Substance:
0/Cations; 0/Detergents; 0/Myelin Proteins

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


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