title: Short peptide motifs for long-lasting anchoring to the cell surface.
abstract: A rational design strategy has been developed for the construction of stable peptide-based anchors for the efficient modification of cell surfaces. Six types of peptide composed of five residues with divalent hydrophobic groups have been designed using this new strategy. Among them, a peptide with a sequence of NBD-Lys-Lys(X)-Lys-Lys-Lys(X)-NH2 (NBD: fluorophore, Lys(X): N-ε-palmitoyl-l-lysine) was found to show the highest modification efficacy and longevity in culture medium. The good performance of this peptide was attributed to (1) its high aqueous solubility, which allowed it to partition from the medium to the cell surface, and (2) the high binding affinity of the saturated palmitoyl groups to the cell membrane. We found that the distribution of the peptide was affected by recycling endosome, which enabled the representation of the peptide following its endocytotic disappearance from the cell membrane. Biotin was also presented on the cell surface using this peptide-based anchor to examine its recognition by streptavidin. The efficacy of the recognition process increased as the length of the oligoethylene glycol spacer increased, indicating that it was necessary for the biotin tag to move away from the membrane glycoproteins on the cell surface to facilitate its efficient recognition by streptavidin.
authors.affiliation: Graduate School of Systems Life Science, ‡Department of Applied Chemistry, §Center for Future Chemistry, and ∥Center for Molecular Systems, Kyushu University , 744 Moto-oka, Nishi-ku, Fukuoka 819-0395, Japan.
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Matsuda
authors.forename: Wataru
authors.initials: W
authors.lastname: Hatanaka
authors.forename: Masafumi
authors.initials: M
authors.lastname: Takeo
authors.forename: Chan Woo
authors.initials: CW
authors.lastname: Kim
authors.forename: Takuro
authors.initials: T
authors.lastname: Niidome
authors.forename: Tatsuhiro
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Akihiro
authors.initials: A
authors.lastname: Kishimura
authors.forename: Takeshi
authors.initials: T
authors.lastname: Mori
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Katayama
mesh_terms: D020816:Amino Acid Motifs; D001665:Binding Sites; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D020014:K562 Cells / Q000187:drug effects / Q000378:metabolism; D008024:Ligands; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D015202:Protein Engineering / Q000379:methods; D019809:Streptavidin / Q000737:chemistry
chemical_list: D008024:Ligands; D010455:Peptides; D001710:Biotin; D019809:Streptavidin