Peptide microarrays for profiling of modification state-specific antibodies.
The reversible phosphorylation of serine, threonine, and tyrosine residues is one of the most important intracellular post-translational modifications regulating enzymatic activities and protein/protein interaction in eukaryotic cells. Tools for determining phosphorylation status of proteins and peptides play a prominent role in signal transduction research and proteomics. Pan-specific antibodies claimed to recognize modified amino acid residues independent on the nature of surrounding residues in peptides and proteins are widely used. We used high-content phosphopeptide microarrays and microarrays displaying acetyllysine-containing peptides for comprehensive characterization of commercially available generic anti-phosphopeptide and anti-acetyllysine antibodies. We were able to demonstrate distinct subsite specificity and cross-reactivity for such antibodies.
JPT Peptide Technologies GmbH, Volmerstr. 5, D-12489 Berlin, Germany.
Johannes
J
Zerweck
Antonia
A
Masch
Mike
M
Schutkowski
D000906:Antibodies / Q000032:analysis* / Q000276:immunology*; D001666:Binding Sites, Antibody; D003429:Cross Reactions; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000276:immunology; D010455:Peptides / Q000737:chemistry* / Q000276:immunology*; D010766:Phosphorylation; D040081:Protein Array Analysis / Q000379:methods*
D000906:Antibodies; D010455:Peptides; C002315:N(alpha)-acetyllysine; D008239:Lysine