Question: 2. It has been established that the so-called ‘ ZZ domain’, a synthetic IgG binding protein deriv…

2. It has been established that the so-called ‘ ZZ
domain’, a synthetic IgG binding protein derived
from tandem repeats

of the B domain of protein A, interacts with the Fc
portion of IgG molecules. HOW would you confirm that this ‘ ZZ
domain’ indeed binds to the Fc region of IgG and not to that of IgM
or IgE

a.What would you establish as criteria
for physiologically relevant binding of this ZZ domain to IgG Fc
portion?

b.What would you use as ‘ligand’ to
determine binding to the ‘ receptor’ , in this case
IgG

c.How would you assure the specificity
of the ZZ domain for the Fc portion of IgG and not that of other
immunoglobulins?

3. The signaling cascade of the FAS receptor, a first
step in the extrinsic apoptotic pathway, includes a variety of
critical protein-protein interactions

a.How would you isolate the protein
that interacts with the DD, or Death Domain, of the FAS
R?

b.How would you modify the cDNA
encoding this DD to identify other interacting proteins in
subpopulations of lymphocytes that interacted with the FAS R in Fas
ligand-stimulated cells, but NOT in resting, or unstimulated,
lymphocyte cells.

c.How would you test the SPECIFICITY of
the Fas R DD domain interactions with your identified interacting
proteins to be sure they uniquely interact with the DD
domain amino acid sequence?