Mapping protein-protein interactions by affinity-directed mass spectrometry.

AUTOR(ES)

Zhao, Y

RESUMO

A precise and rapid method for identifying sites of interaction between proteins was demonstrated; the basis of the method is direct mass spectrometric readout from the complex to determine the specific components of the proteins that interact--a method termed affinity-directed mass spectrometry. The strategy was used to define the region of interaction of a protein growth factor with a monoclonal antibody. A combination of proteolytic digestion and affinity-directed mass spectrometry was used to rapidly determine the approximate location of a continuous binding epitope within the growth factor. The precise boundaries of the binding epitope were determined by affinity-directed mass spectrometric analysis of sets of synthetic peptide ladders that span the approximate binding region. In addition to the mapping of such linear epitopes, affinity-directed mass spectrometry can be applied to the mapping of other types of molecule-molecule contacts, including ligand-receptor and protein-oligonucleotide interactions.

Documentos Relacionados

• Protein sequencing by tandem mass spectrometry.
• Mapping of protein-protein interactions within the DNA-dependent protein kinase complex.
• Control of translational repression by protein-protein interactions.
• Principles of protein-protein interactions.
• Anchor residues in protein–protein interactions