Ligand Screen Experiments
The Ligand Screen is a strategy for detecting the inputs and, in
time, the combinations of inputs that are most relevant to regulation
of the behavior of the cells chosen as our experimental targets. The
initial goals are (1) to determine which ligands give functionally
unique responses and (2) to determine which combinations of ligands
interact in ways that are not simply energetically
additive. Definition of the extent of the interactions among ligands
is a significant goal. The combinations of inputs that display the
most robust interactions will be those to be analyzed rigorously in
later stages of the Alliance's life, since these interactions define
the level of complexity of the signaling network.
We are using a modest number of information-rich assays to identify
unique inputs and interactive inputs. The assays have been chosen to
sample broadly the signaling capability of the cell and the complexity
of the response. Measurements for this phase of experimentation have
not been chosen to yield critical mechanistic information. Assays will
include mRNA profiles, immunoblots to detect phosphorylation at
specific sites within a panel of signaling proteins, and
time-dependent changes in concentrations of cyclic AMP and Ca2+.
Other assays will be added in the future.
The ligand names in the first column are linked to descriptions of the
ligands and the entries in the table are linked to the data for that
ligand/assay combination.
instructions
Data archives (cAMP, calcium, phosphoprotein, etc.)
*In preliminary tests, multiple doses of these
ligands were tested and no measureable increases in intracellular cAMP
were observed. The effects of these ligands on cAMP were therefore
not characterized further in the single ligand screen
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