Document Detail


Synthesis of biotin-tagged chemical cross-linkers and their applications for mass spectrometry.
MedLine Citation:
PMID:  19412923     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Chemical cross-linking combined with mass spectrometry (MS) has been used to elucidate protein structures and protein-protein interactions. However, heterogeneity of the samples and the relatively low abundance of cross-linked peptides make this approach challenging. As an effort to overcome this hurdle, we have synthesized lysine-reactive homobifunctional cross-linkers with the biotin in the middle of the linker and used them to enrich cross-linked peptides. The reaction of biotin-tagged cross-linkers with purified HIV-1 CA resulted in the formation of hanging and intramolecular cross-links. The peptides modified with biotinylated cross-linkers were effectively enriched and recovered using a streptavidin-coated plate and MS-friendly buffers. The enrichment of modified peptides and removal of the dominantly unmodified peptides simplify mass spectra and their analyses. The combination of the high mass accuracy of Fourier transform ion cyclotron resonance (FT-ICR) MS and the tandem mass spectrometric (MS/MS) capability of the linear ion trap allows us to unambiguously identify the cross-linking sites and additional modification, such as oxidation.
Authors:
Sebyung Kang; Liyuan Mou; Jason Lanman; Sadanandan Velu; Wayne J Brouillette; Peter E Prevelige
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Rapid communications in mass spectrometry : RCM     Volume:  23     ISSN:  1097-0231     ISO Abbreviation:  Rapid Commun. Mass Spectrom.     Publication Date:  2009 Jun 
Date Detail:
Created Date:  2009-05-18     Completed Date:  2009-08-11     Revised Date:  2011-09-26    
Medline Journal Info:
Nlm Unique ID:  8802365     Medline TA:  Rapid Commun Mass Spectrom     Country:  England    
Other Details:
Languages:  eng     Pagination:  1719-26     Citation Subset:  IM    
Copyright Information:
Copyright (c) 2009 John Wiley & Sons, Ltd.
Affiliation:
Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35294-2170, USA.
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MeSH Terms
Descriptor/Qualifier:
Biotin / chemical synthesis,  chemistry*
Capsid Proteins / chemistry,  metabolism
Cross-Linking Reagents / chemical synthesis,  chemistry*
HIV-1 / chemistry,  metabolism
Spectroscopy, Fourier Transform Infrared
Tandem Mass Spectrometry / methods*
Grant Support
ID/Acronym/Agency:
AI44626/AI/NIAID NIH HHS; AI53821/AI/NIAID NIH HHS; R01 AI044626-08/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Capsid Proteins; 0/Cross-Linking Reagents; 58-85-5/Biotin
Comments/Corrections

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