Document Detail


Simple approach to assign disulfide connectivity using extracted ion chromatograms of electron transfer dissociation spectra.
MedLine Citation:
PMID:  23210856     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Increasing interest in production of protein-based pharmaceuticals (biotherapeutics) is accompanied by an increased need for verification of protein folding and correct disulfide bonding. Recombinant protein expression may produce aberrant disulfide bonds and could result in safety concerns or decreased efficacy. Thus, the thorough analysis of disulfide bonding is a necessity for protein therapeutics. The use of electron transfer dissociation (ETD) facilitates this analysis because disulfide bonds are preferentially cleaved when subjected to ETD. Here, we make use of this well-characterized reaction to assign disulfide bonding networks by coupling the use of extracted ion chromatograms (XICs) of cysteine-containing peptides with ETD analysis to produce an efficient assignment approach for disulfide bonding. This method can be used to assign a disulfide pattern in a de novo fashion, to detect disulfide shuffling, and to provide information on heterogeneity, when more than one disulfide bonding pattern is present. The method was applied for assigning the disulfide-bonding network of a recombinant monomer of the HIV envelope protein gp120. It was found that one region of the protein, the V1/V2 loops, had significant heterogeneity in the disulfide bonds.
Authors:
Daniel F Clark; Eden P Go; Heather Desaire
Related Documents :
21687866 - Template-free synthesis and characterization of mesoporous tungsten nitride nanoplates.
12688756 - A new type of c2-symmetric bisphospine ligands with a cyclobutane backbone: practical s...
17165946 - Catalytic and highly enantioselective friedel-crafts alkylation of aromatic ethers with...
25434626 - Mn(ii) complexes of scorpiand-like ligands. a model for the mnsod active centre with hi...
21828596 - A kinetic analysis on non-isothermal glass-crystal transformation in ge(1-x)sn(x)se(2.5...
23261796 - Thermally induced isomerization of linoleic acid in soybean oil.
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2013-01-03
Journal Detail:
Title:  Analytical chemistry     Volume:  85     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2013 Jan 
Date Detail:
Created Date:  2013-01-15     Completed Date:  2013-11-26     Revised Date:  2014-01-23    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1192-9     Citation Subset:  IM    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Cysteine / chemistry
Disulfides / chemistry*
Electron Transport
HIV Envelope Protein gp120 / chemistry*
Ions / chemistry
Peptides / chemistry
Recombinant Proteins / chemistry
Tandem Mass Spectrometry
Grant Support
ID/Acronym/Agency:
R01 AI094797/AI/NIAID NIH HHS; R01 GM103547/GM/NIGMS NIH HHS; R01 RR026061/RR/NCRR NIH HHS; R01AI094797/AI/NIAID NIH HHS; R01GM103547/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Disulfides; 0/HIV Envelope Protein gp120; 0/Ions; 0/Peptides; 0/Recombinant Proteins; K848JZ4886/Cysteine
Comments/Corrections

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


Previous Document:  Transcriptionally active heterotrophic diazotrophs are widespread in the upper water column of the A...
Next Document:  Three-carbon Homologation of Diorganozincs with Lithiated Acetylenic Epoxides.