SPOTing Acetyl-Lysine Dependent Interactions.
Post translational modifications have been recognized as chemical signals that create docking sites for evolutionary conserved effector modules, allowing for signal integration within large networks of interactions. Lysine acetylation in particular has attracted attention as a regulatory modification, affecting chromatin structure and linking to transcriptional activation. Advances in peptide array technologies have facilitated the study of acetyl-lysine-containing linear motifs interacting with the evolutionary conserved bromodomain module, which specifically recognizes and binds to acetylated sequences in histones and other proteins. Here we summarize recent work employing SPOT peptide technology to identify acetyl-lysine dependent interactions and document the protocols adapted in our lab, as well as our efforts to characterize such bromodomain-histone interactions. Our results highlight the versatility of SPOT methods and establish an affordable tool for rapid access to potential protein/modified-peptide interactions involving lysine acetylation.
Structural Genomics Consortium, Nuffield Department of Medicine, Oxford University, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK,. sarah.picaud@sgc.ox.ac.uk.
Sarah
S
Picaud
Structural Genomics Consortium, Nuffield Department of Medicine, Oxford University, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK,. panagis.filippakopoulos@sgc.ox.ac.uk.|Ludwig Institute for Cancer Research, Nuffield Department of Medicine, Oxford University, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK. panagis.filippakopoulos@sgc.ox.ac.uk.
Panagis
P
Filippakopoulos
SPOT assay; bromodomain; epigenetic readout; lysine acetylation; recognition motif