Protein surface recognition by synthetic agents: design and structural requirements of a family of artificial receptors that bind to cytochrome c.
The design, synthesis, and evaluation of a novel series of receptors for protein surface recognition are described. The design of these agents is based around the attachment of four constrained peptide loops onto a central calix[4]arene scaffold. This arrangement mimics the role of the hypervariable loops in antibody combining regions and defines a large surface area for binding to a complementary region of the exterior of a target protein. Using affinity and gel filtration chromatographies we show that one particular receptor binds strongly to the surface of cytochrome c. The site of binding is presumably close to the heme edge region, which contains several charged lysine residues. This is supported by the observation that the receptor inhibits the reduction of Fe(III) cytochrome c to its Fe(II) form. We also show that binding is strongly dependent on the nature of the substituents on the lower rim of the calixarene. The nmr and computational studies suggest that this effect may be due to conformational differences among the differently substituted receptors.
Lin
Q
Q
Department of Chemistry, Yale University, New Haven, CT 06511, USA.
Park
H S
HS
Hamuro
Y
Y
Lee
C S
CS
Hamilton
A D
AD
D000818:Animals; D000906:Antibodies / Q000737:chemistry; D001205:Ascorbic Acid / Q000378:metabolism; D001665:Binding Sites; D047250:Calixarenes; D002850:Chromatography, Gel; D003574:Cytochrome c Group / Q000378:metabolism*; D006736:Horses; D007700:Kinetics; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008958:Models, Molecular; D008968:Molecular Conformation; D015394:Molecular Structure; D010084:Oxidation-Reduction; D010456:Peptides, Cyclic / Q000138:chemical synthesis; D011485:Protein Binding; D015202:Protein Engineering; D011956:Receptors, Cell Surface / Q000737:chemistry*
D000906:Antibodies; D003574:Cytochrome c Group; D046911:Macromolecular Substances; D010456:Peptides, Cyclic; D011956:Receptors, Cell Surface; D047250:Calixarenes; D001205:Ascorbic Acid