PROTEIN PAINTING REVEALS HIDDEN PROTEIN-PROTEIN INTERACTION DOMAINS


Year of Award:
2013
Award Type:
R21
Project Number:
CA177535
RFA Number:
RFA-CA-12-002
Technology Track:
Molecular & Cellular Analysis Technologies
PI/Project Leader:
LIOTTA, LANCE ALLEN
Other PI or Project Leader:
N/A
Institution:
GEORGE MASON UNIVERSITY
We propose the creation of a wholly novel technology 'protein painting', for the rapid, direct isolation and sequencing of hidden native protein-proteininteraction domains. Protein-protein interactions are the basis of virtually all functional moleculr events driving cancer cell signaling and gene regulation. Currently no technology exists to directly isolate and sequence unknown interaction domains, or to directly detect whether known protein-protein binding domains are in contact in a cellular or tissue sample of native proteins. Tracking the interface domains between interacting proteins, or within misfolded proteins, is the basis for the next generation of therapies that block the molecular interactions driving cancer. We will create protein paint chemistries, a novel panel of synthetic organic small molecules that bind to protein molecules with high affinity and mask all the protease cleavage sites of the protein. We will use our new protein painting chemistry to isolate and sequence protein-protein interaction domains of cancer related proteins. Interacting native proteins in solution are paintedwith a palette of paint molecules that coat the exposed surfaces of the proteins but do not have access to the internal protein-protein contact domains. Thus if two native proteins are bound together, the interface domains will remain non-painted. The protein paints coat with a high resolution (