Identification of a peptide binding motif for secreted frizzled-related protein-1.
Secreted Frizzled-related proteins (sFRPs) bind Wnts and modulate their activity. To identify putative sFRP-1 binding motifs, we screened an M13 phage displayed combinatorial peptide library. A predominant motif, L/V-VDGRW-L/V, was present in approximately 70% of the phage that bound sFRP-1. Use of peptide/alkaline phosphatase chimeras and alanine scanning confirmed that the conserved motif was important for sFRP-1 recognition. The dissociation constant for a peptide/sFRP-1 complex was 3.9 microM. Additional analysis revealed that DGR was the core of the binding motif. Although Wnt proteins lack this sequence, other proteins possessing the DGR motif may function as novel binding partners for sFRP-1.
Chuman
Yoshiro
Y
Laboratory of Cellular and Molecular Biology, National Cancer Institute, Bethesda, MD 20892-4256, USA.
Uren
Aykut
A
Cahill
Jennifer
J
Regan
Carole
C
Wolf
Vladimir
V
Kay
Brian K
BK
Rubin
Jeffrey S
JS
D000409:Alanine / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D017104:Bacteriophage M13 / Q000737:chemistry / Q000235:genetics; D001667:Binding, Competitive; D002151:Calorimetry; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D051153:Wnt Proteins; D000099274:Frizzled-Related Proteins
D000409:Alanine; D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D011506:Proteins; D011994:Recombinant Proteins; D051153:Wnt Proteins; D000099274:Frizzled-Related Proteins