Journal
JOURNAL OF PROTEOME RESEARCH
Volume 9, Issue 12, Pages 6467-6478Publisher
AMER CHEMICAL SOC
DOI: 10.1021/pr100707t
Keywords
peptide microarray; chemical modification; desorption/ionization; silicon nanowires; mass spectrometry; Mycobacteria; heparin-binding hemagglutinin (HBHA); lysine; methylation; reductive alkylation
Categories
Funding
- CNRS
- Universite de Lille Nord de France
- Institut Pasteur de Lille
- [IFR142]
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Peptide microarrays are useful tools for the characterization of humoral responses against peptide antigens The study of post-translational modifications requires the printing of appropriately modified peptides, whose synthesis can be time-consuming and expensive We describe here a method named chips from chips, which allows probing the presence of antibodies directed toward modified peptide antigens starting from unmodified peptide microarrays The chip from chip concept is based on the modification of peptide microspots by simple chemical reactions The starting peptide chip (parent chip) is covered by the reagent solution, thereby allowing the modification of specific residues to occur, resulting in the production of a modified peptide chip (daughter chip) Both parent and daughter chips can then be used for interaction studies The method is illustrated using reductive methylation for converting lysines into dimethyllysines The rate of methylation was studied using specific antibodies and fluorescence detection, or surface-assisted laser desorption ionization mass spectrometry This later technique showed unambiguously the efficient methylation of the peptide probes The method was then used to study the humoral response against the Mycobacterium tuberculosis heparin-binding hemagglutinin, a methylated surface-associated virulence factor and powerful diagnostic and protective antigen
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