Peptides that mimic glycosaminoglycans: high-affinity ligands for a hyaluronan binding domain.
BACKGROUND
Hyaluronan (HA) is a non-sulfated glycosaminoglycan (GAG) that promotes motility, adhesion, and proliferation in mammalian cells, as mediated by cell-surface HA receptors. We sought to identify non-carbohydrate ligands that would bind to and activate cell-surface HA receptors. Such analogs could have important therapeutic uses in the treatment of cancer, wound healing, and arthritis, since such ligands would be resistant to degradation by hyaluronidase (HAse).


RESULTS
Peptide ligands that bind specifically to the recombinant HA binding domain (BD) of the receptor for hyaluronan-mediated motility (RHAMM) were obtained by screening two peptide libraries: (i) random 8-mers and (ii) biased 8-mers with alternating acidic side chains, i.e. XZXZXZXZ (X=all-L-amino acids except Cys, Lys, or Arg; Z=D-Asp, L-Asp, D-Glu, or L-Glu). Selectivity of the peptide ligands for the HABD was established by (i) detection of binding of biotin- or fluorescein-labeled peptides to immobilized proteins and (ii) fluorescence polarization of FITC-labeled peptides with the HABD in solution. HA competitively displaced binding of peptides to the HABD, while other GAGs were less effective competitors. The stereochemistry of four biased octapeptides was established by synthesis of the 16 stereoisomers of each peptide. Binding assays demonstrated a strong preference for alternating D and L configurations for the acidic residues, consistent with the calculated orientation of glucuronic acid moieties of HA.


CONCLUSIONS
Two classes of HAse-resistant peptide mimetics of HA were identified with high affinity, HA-compatible binding to the RHAMM HABD. This demonstrated that non-HA ligands specific to a given HA binding protein could be engineered, permitting receptor-specific targeting.
Ziebell
M R
MR
Department of Physiology and Biophysics, The University at Stony Brook, NY 11794-8661, USA.
Zhao
Z G
ZG
Luo
B
B
Luo
Y
Y
Turley
E A
EA
Prestwich
G D
GD
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D016326:Extracellular Matrix Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D006025:Glycosaminoglycans / Q000037:antagonists & inhibitors* / Q000737:chemistry; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D006820:Hyaluronic Acid / Q000037:antagonists & inhibitors / Q000737:chemistry*; D008024:Ligands; D008958:Models, Molecular; D018716:Molecular Mimicry; D009842:Oligopeptides / Q000737:chemistry* / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D013329:Structure-Activity Relationship
D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D018960:Hyaluronan Receptors; D008024:Ligands; D009842:Oligopeptides; D019151:Peptide Library; C098946:hyaluronan-mediated motility receptor; D006820:Hyaluronic Acid