AfCS Yeast 2-Hybrid Interaction Data
Identification of protein-protein interactions and the changes in
such interactions that result from ligand binding or covalent
modification are clearly central to understanding the mechanisms of
signal transduction and establishing intracellular signaling networks.
The AfCS is utilizing high-throughput methods to detect
protein-protein interactions between signaling molecules expressed in
B cells and cardiac myocytes. The AfCS has arranged a collaboration
with Myriad
Genetics to perform large-scale yeast two-hybrid screens.
The yeast two-hybrid system is a well-established methodology that
permits the rapid identification of binary interactions between a
chosen test protein (the "bait") and an interacting protein (the
"prey"). This is achieved by expressing the bait protein as a hybrid,
fused to the DNA binding domain of a transcription factor, and
screening it against a library of prey candidates that are fused to
the corresponding transcriptional activation domain. Both fusions are
expressed in yeast cells that carry a reporter gene whose expression
is under the control of the transcription factor, such that the
interaction of the "two hybrids" leads to expression of the
reporter.
The AfCS System Committees selected B cell and myocyte signaling
proteins that were considered optimal candidates for yeast two-hybrid
screens. Myriad has developed an industrial-scale system, termed ProNet®, for the discovery of
protein-protein interactions via the yeast two-hybrid system. An
efficient mating protocol, which uses proprietary strains and vectors,
was used to search AfCS-selected baits against different AfCS
activation domain libraries.
Return to yeast two-hybrid display
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