Biomedical Engineering Reference
In-Depth Information
A chemical proteomics approach was used to identify the target protein of
Wntepane 1. A set of biotin-tagged bait compounds, including an affinity probe
(Figure 18.9), was synthesized and immobilized on streptavidin-coated magnetic
beads. After treatment with HEK293 cell lysate, the proteins bound to the beads
were eluted with nonbiotinylated Wntepane 1 and separated by polyacrylamide gel
electrophoresis (PAGE). Mass spectrometry identified the van Gogh-like receptor
protein 1 (Vangl1) as the most plausible target. The identification of Vangl1 as the
target of the Wntepanes is notable because Vangl1 works primarily on the noncanoni-
cal Wnt/PCP pathway [111]. Interestingly, the activators targeting Vangl1 were found
work synergistically with the canonical Wnt ligand Wnt3a.
As the first activators of Wnt signaling that target Vangl1 that have been identified,
the Wntepanes can serve as novel probes for studying the activation of the Wnt path-
ways through Vangl1, the relationship between the noncanonical Wnt/PCP pathway
and the canonical Wnt pathway, and Wnt pathway protein activities in cancer and
stem cell biology.
18.7 SUMMARY AND OUTLOOK
DOS strategies have greatly facilitated the discovery of new and important biologi-
cal probes in the past decade. This success has yielded new insight into complex
biological processes and afforded leads for developing novel therapies. We can
expect that the potential of the DOS approach to probe development will con-
tinue to emerge in future studies, especially studies aimed at developing high-
quality probes that affect a challenging protein target or biological process [112].
An overarching aim of the DOS approach to probe development is to provide a
bridge between basic biology and human diseases. This aim will become a reality
when a broad range of probes has become accessible to the research and medical
communities.
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