 |
PDBsum entry 3svv
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Transferase/transferase inhibitor
|
PDB id
|
|
|
|
3svv
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
References listed in PDB file
|
|
|
Key reference
|
|
|
Title
|
|
Chemical genetic strategy for targeting protein kinases based on covalent complementarity.
|
|
|
Authors
|
|
A.L.Garske,
U.Peters,
A.T.Cortesi,
J.L.Perez,
K.M.Shokat.
|
|
|
Ref.
|
|
Proc Natl Acad Sci U S A, 2011,
108,
15046-15052.
|
|
|
PubMed id
|
|
|
|
|
Note: In the PDB file this reference is
annotated as "TO BE PUBLISHED". The citation details given above have
been manually determined.
|
|
|
|
Abstract
|
|
|
The conserved nature of the ATP-binding site of the > 500 human kinases
renders the development of specific inhibitors a challenging task. A widely used
chemical genetic strategy to overcome the specificity challenge exploits a
large-to-small mutation of the gatekeeper residue (a conserved hydrophobic amino
acid) and the use of a bulky inhibitor to achieve specificity via shape
complementarity. However, in a number of cases, introduction of a glycine or
alanine gatekeeper results in diminished kinase activity and ATP affinity. A new
chemical genetic approach based on covalent complementarity between an
engineered gatekeeper cysteine and an electrophilic inhibitor was developed to
address these challenges. This strategy was evaluated with Src, a
proto-oncogenic tyrosine kinase known to lose some enzymatic activity using the
shape complementarity chemical genetic strategy. We found that Src with a
cysteine gatekeeper recapitulates wild type activity and can be irreversibly
inhibited both in vitro and in cells. A cocrystal structure of T338C c-Src with
a vinylsulfonamide-derivatized pyrazolopyrimidine inhibitor was solved to
elucidate the inhibitor binding mode. A panel of electrophilic inhibitors was
analyzed against 307 kinases and MOK (MAPK/MAK/MRK overlapping kinase), one of
only two human kinases known to have an endogenous cysteine gatekeeper. This
analysis revealed remarkably few off-targets, making these compounds the most
selective chemical genetic inhibitors reported to date. Protein engineering
studies demonstrated that it is possible to increase inhibitor potency through
secondary-site mutations. These results suggest that chemical genetic strategies
based on covalent complementarity should be widely applicable to the study of
protein kinases.
|
|
|
|
|
|
|
