Fig. 7. (A) Binding of synthetic peptides (see
Fig. 6A) to purified GST-fusion
proteins of paxillin (expected Mr approx. 95 kDa) and of
the N-terminal domain (amino acids 1-407) of human Pyk2 (expected
Mr approx. 70 kDa). Bound and unbound proteins were
analyzed by immunoblotting with an anti-GST antibody as well as on
Coomassie-stained 15% SDS-polyacrylamide gels. Note the depletion of
GST-fusion proteins in the supernatants incubated with the ß3-tail
peptide. Note also the nearly complete absence of bovine serum albumin in the
bound fractions, which was present in a very large excess over GST-fusion
proteins in the binding reactions. (Co, control resin consisting of
streptavidin agarose without any added peptide; n.d., not done). (B)
Comparison of GST, GST-paxillin and GST-Pyk2NT binding to the ß3-tail
peptide. Note the depletion in the unbound fractions of GST-Pyk2NT and
GST-paxillin after binding. GST proteins were calibrated to give comparable
signals in the supernatants by detection with Anti-GST antibody.
