Methods for identifying protein-protein interactions

摘要

The invention generally relates to methods for identifying protein-protein interactions. In certain aspects, methods of the invention involve conducting a protein-fragment complementation assay on a sample to form a protein-protein complex between two proteins in the sample that only transiently interact under physiological conditions, separating the complexes from the sample, and analyzing a protein of the complex using a mass spectrometry technique.

主权项

1. A method for analyzing two transiently interacting proteins in a sample, wherein a first protein is fused with a first fragment of a reporter protein and a second protein is fused with a second fragment of said reporter protein, the method comprising:
conducting a protein-fragment complementation assay on the sample to form a protein-protein complex transiently under physiological conditions; separating the complex from the sample using a moiety that specifically binds a fused form of the reporter protein but not either of the first and second proteins individually, and applying a washing condition to the sample that removes the first and second proteins in their free form while preserving the complex; and quantitatively analyzing at least one protein of the complex using a mass spectrometry technique.