TY - JOUR
T1 - Peptides that mimic glycosaminoglycans
T2 - High-affinity ligands for a hyaluronan binding domain
AU - Ziebell, Michael R.
AU - Zhao, Zhan Gong
AU - Luo, Bai
AU - Luo, Yi
AU - Turley, Eva A.
AU - Prestwich, Glenn D.
N1 - Funding Information:
This work was supported by grants to G.D.P. from the US Department of Defense, the State of Utah Centers of Excellence Program, and The University of Utah (UT, USA), and by a grant to E.A.T. from the National Cancer Institute of Canada. We thank Dr. J.T. Elliott (The University at Stony Brook, USB) and Professor R. El Magrahbi (USB) for biophysical advice, Mr. R. Gerhart (UT, USA) for bead-picking, and Dr. R.W. Schackmann (UT, USA) for peptide sequencing and synthesis.
PY - 2001
Y1 - 2001
N2 - 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-competable 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.
AB - 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-competable 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.
KW - Glycosaminoglycan-mimicking peptide
KW - High-affinity ligand
KW - Hyaluronan binding domain
KW - Receptor for hyaluronan-mediated motility
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U2 - 10.1016/S1074-5521(01)00078-3
DO - 10.1016/S1074-5521(01)00078-3
M3 - Article
C2 - 11731299
AN - SCOPUS:0035202460
SN - 1074-5521
VL - 8
SP - 1081
EP - 1094
JO - Chemistry and Biology
JF - Chemistry and Biology
IS - 11
ER -