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A chemically synthesized analog of Bacillus cereus secondary cell wall polysaccharide (SCWP) facilitates the identification of PatB1 as a SCWP O-acetyltransferase, and the structure of PatB1 provides insights into its catalytic mechanism.
The small molecule nitazoxanide (NTZ) was identified as a Wnt inhibitor by promoting protein citrullination of β-catenin through increased cytosolic calcium and PAD2 protein stabilization. β-catenin citrullination results in proteasomal degradation.
N-Acetylglucosamine derivatives equipped with electrophilic groups and handles for subsequent chemical tagging are useful probes of substrate recognition by O-GlcNAc transferases and enable the capture of transient protein substrates of these enzymes.
The development of a computational protein design method, meta-multistate design, enables the design and validation of protein variants termed DANCERs that spontaneously exchange between predicted conformational states on the millisecond timescale.
A positive-selection CRISPR screen with the pro-oxidant paraquat (PQ) uncovers three genes mediating PQ-induced cell death: POR is the source of PQ-mediated reactive oxygen species (ROS) generation, and ATP7A and SLC45A4 promote oxidant-dependent cell death.
The use of phage-assisted continuous evolution (PACE) with both positive and negative selection enables the rapid development of orthogonal aminoacyl-tRNA synthetases with high activity and selectivity for noncanonical amino acids.
Microtubule sliding driven by kinesin-14 HSET is regulated by a feedback mechanism. When microtubules start sliding apart, HSET molecules are retained in the shortening overlap, which leads to an HSET-density-dependent decrease in sliding velocity.
The N-terminal domain structure of pyrrolysyl-tRNA synthetase (PylRS) reveals details of its tRNA specificity and facilitates the improvement of its selectivity for non-canonical amino acids by phage-assisted non-continuous evolution (PANCE).
A linear gating mechanism links kinetically and structurally distinct closed and open states of NMDA receptors. During allosteric inhibition, agonist binding incudes uncoupling of structural changes from gating motions in the first transmembrane region.
A systems chemical biology approach to characterize beneficial off-target effects revealed a polypharmacology mechanism for the multikinase inhibitor ceritinib and a repurposing opportunity through rational design of a synergistic drug combination.
The natural product stendomycin is a first-in-class inhibitor of the TIM23 mitochondrial translocon in yeast and mammalian cells that helped reveal that the TIM23 complex does not regulate transport of the autophagy regulator PINK1.
An inhibitor of the deubiquitinase (DUB) USP10 regulates the degradation of oncogenic FLT3, thus defining USP10 as a DUB for FLT3 and providing a therapeutic approach for human acute myeloid leukemia in which FLT3 activation is dysregulated.
Kinetic analysis and EPR spectroscopy measurements on a bacterial heme-nitric oxide/oxygen-binding (H-NOX) protein reveal that nitric oxide (NO) binds on the distal side of the heme cofactor under both limiting and excess NO conditions.
The crystal structure of the fluorogenic RNA aptamer Corn reveals an unexpected homodimer. One molecule of the cognate fluorophore DFHO is encapsulated at the locally asymmetric RNA interface, which is notable for lacking intermolecular base pairs.
A highly photostable RNA mimic of red fluorescent protein, Corn, was designed and used to image RNA polymerase III transcription. Quantitative imaging of Corn-tagged transcripts revealed transcription dynamics in live cells following mTOR inhibition.
Single-molecule FRET microscopy and hidden Markov modeling analysis with the full-length SAM-I riboswitch reveal the existence of multiple conformational states and the effects of Mg2+ and SAM binding on their dynamics and population shifts.
A conserved negative regulator of TORC1, GATOR1, regulates the synthesis of the nitrogen-containing amino acids aspartate and glutamine from the mitochondrial TCA cycle in tune with the amino acid and nitrogen status of cells.
An inhibitor of the SUMO-activating (E1) enzyme SAE blocks enzyme activity and total SUMOylation in cells—thereby defining roles for SAE in mitotic progression and chromosome segregation—and also decreases cancer cell proliferation.
In Pseudoalteromonas rubra, an unclustered biosynthetic gene encodes a di-iron oxygenase-like enzyme that catalyzes regiospecific C–H activation and cyclization of prodigiosin, yet is unrelated to the Rieske oxygenases that produce other cyclized prodiginines.
Engineering of an artificial metabolon complex consisting of an endogenous sugar transporter and a heterologous isomerase couples the uptake of xylose to its downstream reaction, improving ethanol yield and minimizing an undesired side reaction.