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Most bisubstrate analogs have been designed to mimic the phosphate donor (ATP) and the acceptor components (Ser-, Thr-, or Tyr-containing peptides).
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Bisubstrate analogs have the potential to provide enhanced specificity for protein kinase inhibition and tools to understand catalytic mechanism.
This chapter also includes an in-depth discussion of several bisubstrate analogs that are currently in use as potent therapeutics.
The use of bisubstrate analogs as inhibitors for enzymatic reactions has proven to be a very effective strategy.
These bisubstrate analogs exhibit stronger potency in the inhibition of Esa1 and Tip60 compared to the small molecules curcumin and anacardic acid.
1.35 Å TAT cocrystal structures with bisubstrate analogs constrain TAT action to the microtubule lumen and reveal Lys40 engaged in a suboptimal active site.
We have also shown that related bisubstrate analogs can be used to potently block serine/threonine kinases including protein kinase A. Since many protein kinases recognize folded protein substrates for efficient phosphorylation, it was advantageous to incorporate the peptide ATP conjugates into protein structures.
Initially, FTIs were designed to be competitive inhibitors, either peptidomimetic compounds, isoprenoid analogs or bisubstrate analogs.
Additional studies on peptide-CoA conjugates have explored the broader potential of these bisubstrate analogs against a range of acetyltransferases.
In contrast, Gcn5 and Pcaf acetyltransferases obey classical ternary complex mechanisms, and are more potently inhibited by the longer peptide-CoA conjugate bisubstrate analogs.
Assays with bisubstrate analogs containing various length linkers between the peptidyl-Lys and the CoA showed that linker length correlated with potency, although these compounds never exceeded the potency of the original Lys-CoA.
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