Screening of alpha-helical peptide ligands controlling a calcineurin-phosphatase activity.
In this paper, we describe an application of 202-membered fluorescently labeled peptide library designed to take an alpha-helix secondary structure. As a proof-of-concept experiment, a calmodulin (CaM)/calcineurin (Cn) pair was chosen to screen alpha-helical peptide ligands that tightly bind to CaM and also control enzymatic functions of Cn. Three peptides were successfully selected from the library by assaying Cn-phosphatase activities and peptide-CaM interactions (dual check process). The strategy using a designed peptide library shows real promise as a peptide-based high-throughput screening system.
Usui
Kenji
K
Department of Bioengineering and The COE21 Program, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, B-40, 4259 Nagatsuta, Midori, Yokohama 226-8501, Japan.
Tomizaki
Kin-Ya
KY
Mihara
Hisakazu
H
D000595:Amino Acid Sequence; D019703:Calcineurin / Q000737:chemistry; D002147:Calmodulin / Q000737:chemistry; D004341:Drug Evaluation / Q000379:methods; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification*; D010744:Phosphoric Monoester Hydrolases / Q000032:analysis*; D017433:Protein Structure, Secondary
D002147:Calmodulin; D008024:Ligands; D019151:Peptide Library; D010455:Peptides; C120817:calcineurin phosphatase; D019703:Calcineurin; D010744:Phosphoric Monoester Hydrolases