Mapping Protein-Protein Interactions by Affinity-Directed Mass Spectrometry
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. Th...
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Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1996-04, Vol.93 (9), p.4020-4024 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | 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. |
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ISSN: | 0027-8424 1091-6490 |
DOI: | 10.1073/pnas.93.9.4020 |