Receptor mediated uptake of peptides that bind the human transferrin receptor

Eur J Biochem. 2001 Apr;268(7):2004-12. doi: 10.1046/j.1432-1327.2001.02073.x.

Abstract

A biopanning process designed to find peptide epitopes specific for cell surface receptors has been used in this study to select seven- and 12-amino-acid peptides capable of binding to and internalizing with the human transferrin receptor (hTfR). Through sequential rounds of negative and positive selection, two peptide sequences were identified that specifically bind to the hTfR. Phage containing the sequences HAIYPRH or THRPPMWSPVWP were inhibited from binding the hTfR in a dose-dependent fashion when peptides of the same sequence were present in a competition assay. Interestingly, transferrin did not compete with either of these sequences for receptor binding, suggesting that these peptides bind a site on the hTfR distinct from the transferrin binding site. When either of these sequences was expressed as a fusion to green fluorescent protein (GFP), the recombinant GFP molecule was internalized in cells expressing the hTfR. These studies suggest that the two peptides can be used to target other proteins into the endosomal pathway. Further, they provide a strategy for identifying peptides that bind to other cell surface receptors that can be used for both diagnostic and therapeutic purposes.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding, Competitive
  • Cell Line
  • Chick Embryo
  • Electrophoresis, Polyacrylamide Gel
  • Endocytosis
  • Fluorescent Antibody Technique
  • Green Fluorescent Proteins
  • Humans
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Molecular Sequence Data
  • Peptides / metabolism*
  • Receptors, Transferrin / metabolism*

Substances

  • Luminescent Proteins
  • Peptides
  • Receptors, Transferrin
  • Green Fluorescent Proteins