title: Development of crosslink-type tag-probe pairs for fluorescent imaging of proteins.
abstract: Useful methods of protein labeling via functional peptide tags have been developed in the field of proteome and chemical biology. New tag-probe pairs based on leucine zipper peptides for labeling target proteins are described. These consist of an α-helical probe peptide with an environmental-sensitive fluorescent dye and two antiparallel α-helical tag peptides, and may be crosslinked, from the Cys residue of the tag peptide to the N(α)-chloroacetyl group of the probe peptide. Binding of the probe peptide to the tag peptides results in movement of the fluorophore from a hydrophilic to a hydrophobic environment inside the leucine zipper structure, causing a dramatic fluorescent change, mediated by the binding of the two peptides. As a spacer between the N(α)-chloroacetyl group and the original probe sequence, a single Gly residue was the most suitable among 0-2 Gly residues. Crosslinking leads to superior fluorescence response, binding affinity, and chemical stability. These ZIP tag-probe pairs are useful for labeling and fluorescent imaging of proteins.
authors.affiliation: Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Chiyoda-ku, Tokyo 101-0062, Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Nomura
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Mino
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Narumi
authors.forename: Nami
authors.initials: N
authors.lastname: Ohashi
authors.forename: Akemi
authors.initials: A
authors.lastname: Masuda
authors.forename: Chie
authors.initials: C
authors.lastname: Hashimoto
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Tsutsumi
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Tamamura
mesh_terms: D005456:Fluorescent Dyes / Q000737:chemistry*; D016350:Leucine Zippers; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*
chemical_list: D005456:Fluorescent Dyes; D015335:Molecular Probes; D010455:Peptides; D011506:Proteins