Covalent inhibitors
have recently seen a resurgence of interest
in drug development. Nevertheless, compounds, which do not rely on
an enzymatic activity, have almost exclusively been developed to target
cysteines. Expanding the scope to other amino acids would be largely
facilitated by the ability to globally monitor their engagement by
covalent inhibitors. Here, we present the use of light-activatable
2,5-disubstituted tetrazoles that allow quantifying 8971 aspartates
and glutamates in the bacterial proteome with excellent selectivity.
Using these probes, we competitively map the binding sites of two
isoxazolium salts and introduce hydrazonyl chlorides as a new class
of carboxylic-acid-directed covalent protein ligands. As the probes
are unreactive prior to activation, they allow global profiling even
in living Gram-positive and Gram-negative bacteria. Taken together,
this method to monitor aspartates and glutamates proteome-wide will
lay the foundation to efficiently develop covalent inhibitors targeting
these amino acids.
G protein-coupled receptors (GPCRs) have been known for decades as attractive drug targets. This has led to the development and approval of many ligands targeting GPCRs. Although ligand binding effects have been studied thoroughly for many GPCRs, there are multiple aspects of GPCR signaling that remain poorly understood. The reasons for this are the difficulties that are encountered upon studying GPCRs, for example, a poor solubility and low expression levels. In this work, we have managed to overcome some of these issues by developing an affinity-based probe for a prototypic GPCR, the adenosine A 1 receptor (A 1 AR). Here, we show the design, synthesis, and biological evaluation of this probe in various biochemical assays, such as SDS-PAGE, confocal microscopy, and chemical proteomics.
Covalent inhibitors have recently seen a
resurgence of interest in drug development. Nevertheless, compounds, that do
not rely on an enzymatic activity, have almost exclusively been developed to
target cysteines. Expanding the scope to other amino acids would be largely
facilitated by the ability to globally monitor their engagement by covalent
inhibitors. Here, we present the use of light-activatable 2,5-disubstituted
tetrazoles that allow quantifying 8971 aspartates and glutamates in the
bacterial proteome with excellent selectivity. Using these probes, we
competitively map the binding sites of two isoxazolium salts and introduce
hydrazonyl chlorides as a new class of carboxylic acid-directed covalent
protein ligands. As the probes are unreactive prior to activation, they allow global
profiling even in living Gram-positive and Gram-negative bacteria. Taken
together, this method to monitor aspartates and glutamates proteome-wide will
lay the foundation to efficiently develop covalent inhibitors targeting these
amino acids
Covalent inhibitors have recently seen a
resurgence of interest in drug development. Nevertheless, compounds, that do
not rely on an enzymatic activity, have almost exclusively been developed to
target cysteines. Expanding the scope to other amino acids would be largely
facilitated by the ability to globally monitor their engagement by covalent
inhibitors. Here, we present the use of light-activatable 2,5-disubstituted
tetrazoles that allow quantifying 8971 aspartates and glutamates in the
bacterial proteome with excellent selectivity. Using these probes, we
competitively map the binding sites of two isoxazolium salts and introduce
hydrazonyl chlorides as a new class of carboxylic acid-directed covalent
protein ligands. As the probes are unreactive prior to activation, they allow global
profiling even in living Gram-positive and Gram-negative bacteria. Taken
together, this method to monitor aspartates and glutamates proteome-wide will
lay the foundation to efficiently develop covalent inhibitors targeting these
amino acids
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.