Utilizing peptide SPOT arrays to identify protein interactions.

SPOT arrays consist of synthesized peptides 12- to 18-amino acids long, with overlapping sequences that cover the entire sequence of a protein, covalently linked to a solid support. This unit describes how to construct peptide SPOT arrays, biotinylate recombinant proteins, and conduct overlay assays to identify binding interactions. In addition, directions describing how to analyze results to determine single amino acid binding contributions are included. The two techniques in this unit describe how to scan protein sequences to find binding motifs and how to conduct site-directed mutagenesis studies. (c) 2008 by John Wiley & Sons, Inc.