IDENTIFICATION OF TARGET-SPECIFIC FOLDING SITES IN PEPTIDES AND PROTEINS

摘要

The invention provides methods for identification and determination of target-specific folding sites in peptides and proteins, including a method for determining a secondary structure binding to a target of interest within a known parent polypeptide that binds to the target of interest. In one embodiment of the invention, a residue or mimetic containing a nitrogen atom and a sulfur atom available for binding to a metal ion is serially substituted for single residues in or inserted between two adjacent residues in a known primary sequence of a peptide or protein. The resulting sequence, which includes a minimum of the residue or mimetic containing a nitrogen atom and a sulfur atom available for binding to a metal ion and two residues on the amino terminus side thereof, is complexed with a metal ion, thereby forming a metallopeptide. The resulting metallopeptides are then used in binding or functional assays related to the target of interest, and the metallopeptide demonstrating binding or functional activity is selected. The invention further provides methods to determine the specific sequence and local three-dimensional structure of that portion of peptides or proteins that bind to a receptor or target of interest, or mediate a biological activity of interest and methods to determine the pharmacophore of receptors or targets of interest. The invention provides for defined pharmacophores or receptors or targets of interest and directed libraries for identification and determination of target-specific folding sites in peptides and proteins and for identification and determination of pharmacophores of receptors or targets of interest.