A general screening strategy for peptide-based fluorogenic ligands: probes for dynamic studies of PDZ domain-mediated interactions.
A systematic and general approach for identifying efficient probes for class I PDZ domains based on environment-sensitive chromophores is presented. A series of peptides derived from the C-terminal sequence of Stargazin was first used with PDZ domains of PSD-95 and Shank3 to identify the optimal position and linker length for the 4-DMAP chromophore. The results were applied to well-characterized ligand sequences for each set of domains to generate high affinity probes that retain their native sequence specificity and yield remarkable fluorescence increases upon binding. These probes constitute efficient tools to study the dynamics and regulatory mechanisms of PDZ domain-mediated interactions.
Sainlos
Matthieu
M
Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139-4307, USA.
Iskenderian
Wendy S
WS
Imperiali
Barbara
B
D015761:4-Aminopyridine / Q000031:analogs & derivatives; D000595:Amino Acid Sequence; D005456:Fluorescent Dyes / Q000737:chemistry / Q000302:isolation & purification*; D008024:Ligands; D008969:Molecular Sequence Data; D054731:PDZ Domains; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification*
D005456:Fluorescent Dyes; D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D015761:4-Aminopyridine; C003885:4-dimethylaminopyridine