Biomedical Engineering Reference
In-Depth Information
with Hap3p-GST fusion protein and visualized using a fluorescently labeled anti-
GST antibody. A true Hap3p binder called haptamide A was revealed in the array
screening. It was later verified by biophysical SPR experiment (
K
d
=
M).
Further SAR analysis revealed a more potent ligand, haptamide B of Hap3p with a
K
d
value of 0.33
5.03
M. Subsequent cell experiments also showed that haptamide A is
a reversible inhibitor for Hap3p-mediated transcription in vivo.
In both aforementioned experiments, chlorinated glass slides were utilized to
capture DOS-derived compounds containing a hydroxyl group. Literature reports,
however, showed that there are more synthetic libraries with carboxylic acids and
phenols than those with a hydroxyl group. Because of this, Schreiber et al. employed
an alternative immobilization strategy to anchor small molecules with a phenol group.
They synthesized a library of 6336-member phenol-containing fused bicycles and
tetracycles with up to six stereogenic centers [19]. The compounds were then printed
(a)
(b)
(c)
FIGURE 13.2
Selected examples of libraries used in SMM and hits identified in the array
experiments.
