Journal
CHEMBIOCHEM
Volume 4, Issue 4, Pages 272-276Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.200390047
Keywords
combinatorial chemistry; fluorescent probes; lanthanides; peptides
Ask authors/readers for more resources
Lanthanide-binding tags (LBTs) are protein fusion partners consisting of encoded amino acids that bind lanthanide ions with high affinity. Herein, we present a new screening methodology for the identification of new LBT sequences with high affinity for Tb3+ ions and intense luminescence properties. This methodology utilizes solid-phase split-and-pool combinatorial peptide synthesis. Orthogonally clevable linkers allow an efficient two-step screening procedure. The initial screen avoids the interference caused by on-bead screening by photochemically releasing a portion of the peptides into an agarose matrix for evaluation. The secondary screen further characterizes each winning sequence in a defined 17 encoded amino acids that demonstrated a 140-fold increase in affinity (57 nM dissociation constant, K-D) over previously reported lanthanide-binding peptides. This linear sequence was macrocyclized by introducing a disulfide bond between flanking cysteine residues to produce a peptide with a 2-nM apparent dissociation constant for Tb3+ ions.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available