Volume 13 Issue 2, February 2017

A phosphodiesterase, CdnP, from Mycobacterium tuberculosis (M. tb.) helps the pathogen evade immune detection by degrading the second messenger cyclic di-AMP that alerts the host to its presence. Genetic knockout of CdnP dampens the virulence of the pathogen, suggesting that CdnP inhibitors are potential anti–M. tb. therapeutics.

Sensing and responding to diverse extracellular signals is a crucial aspect of cellular decision-making that is currently lacking in the synthetic biology toolkit. The development of modular receptor platforms allows for the rewiring of cellular input–output relationships.

The ability to measure the binding of a compound to its intended target in live cells or tissue is a critical parameter for drug discovery. A new method using polarized light microscopy adds to the current toolbox by enabling monitoring of target engagement in vitro and in vivo at single-cell resolution.

Nitrogenase has the canonical ability to reduce N₂ to NH₃, but under certain conditions, either in vitro or in vivo, it has the additional capability to convert CO₂ to CO and CO to light hydrocarbons.

This perspective discusses recent progress in the development of pharmacological tools that initiate mitophagy and spare mitochondrial function and focuses on promising approaches to identify improved reagents.

The iron protein components of bacterial nitrogenases are capable of reducing carbon dioxide (CO₂) to carbon monoxide (CO) in the absence of their catalytic partners, mimicking the activity of CO dehydrogenase.

The development of small-molecule fluorescent probes through addition of a lipidated cysteine residue next to a caged fluorophore enables detection of endogenous cysteine depalmitoylation by acyl–protein thioesterases in vitro and in live cells.

Super-binding peptides based on sequences of glycine receptors that interact with the neuronal scaffold protein gephyrin are useful for isolating and localizing native gephyrin and for modulation of glycinergic synaptic transmission.

Metabolic labeling of the Drosophila proteome using a chemical ligation approach identifies proteins modified by O-GlcNAc transferase (OGT) including Polycomb proteins at homeotic gene loci and at unexpected sites, implicating OGT in gene-expression regulation.

The use of fluorescence-polarized microscopy, combined with competitive binding with matched fluorescence companion imaging probes, enable target engagement measurements of covalent and reversible small molecule inhibitors in a single cell.

Mass-spectrometry-based proteomics led to the identification of NoBody, a microprotein translated from LINC01420 RNA, which interacts with enhancer of decapping 4 (EDC4) and negatively regulates 5′-to-3′ mRNA decay.

Saturation mutagenesis, molecular modeling and biochemical analysis revealed that active site interactions involving Thr1372 of TET2 are responsible for controlling its proficiency for stepwise oxidation of 5-methylcytosine residues within DNA.

Dimeric bis-benzimidazole compounds that bind selectively to toxic expanded r(CUG) RNA repeat sequences were identified and used as a scaffold for covalent modification, site-specific cleavage and on-target assembly of imaging reagents at expanded r(CUG) sequences in cells.

Single-molecule force spectroscopy and thermodynamics studies reveal six conformational states in the formation of a guanine aptamer, with a ‘kissing loop’ playing a key role in the conformational switching.

Synthetic biology enables re-engineering of cellular functions by introduction of modular, orthogonal signaling pathways, as illustrated by the reprogramming of human T cells to produce IL-2 in response to vascular endothelial growth factor (VEGF).

A mycobacterial phosphodiesterase, CdnP, hydrolyzes bacteria-derived 3′,5′-c-di-AMP as well as host-generated 2′,3′-cGAMP, which activates the host cytosolic surveillance pathway, to dampen host responses.

A high-throughput screen identifies inhibitors of the M. tuberculosis dormancy regulation system, DosRST, including compounds that inhibit autophosphorylation of the DosS and DosT sensor kinases and those that inhibit the catalytic heme of these kinases.

Structural and functional characterization of an aromatic prenyltransferase reveals a unique spacious hydrophobic pocket with conformational fluctuation and multiple acceptor binding sites that endow it with uncommon enzymatic promiscuity.

The ‘CoINPocket’ approach identifies pharmacological similarities between G protein–coupled receptors. On the basis of predicted pharmacological similarity to a few phylogenetically unrelated receptors, the approach identified surrogate ligands for the orphan receptor GPR37L1.