Fig. 1.

In vitro binding of wild-type and phospho-mimetic eNOS to GST-Pin1 fusion proteins. GST fusion proteins representing full-length (FL) human Pin1 (residues 1–163), Δ WW Pin1 (residues 43–163), and a GST non-fusion protein were expressed in E. coli and purified by affinity binding to glutathione–Sepharose. Proteins, prebound to beads, were incubated with purified baculovirus-expressed bovine wild-type (WT) and S116D eNOS. Following binding, extensive washing, and boiling in SDS sample buffer, proteins were immunoblotted (IB) with anti-eNOS antibody. A, Representative blot. B, Densitometric analysis of 3 blots from 3 separate experiments (means ± S.E.).