Research articles

Acidification enhances lactate dehydrogenase– and malate dehydrogenase–mediated promiscuous production of L-2-hydroxyglutarate (L-2HG) from α-ketoglutarate and stabilizes HIF-1α levels.

The application of strand-specific total RNA sequencing combined with metagene analysis enables detection of small-molecule-mediated effects on transcription initiation, elongation or RNA processing, and reveals that isoginkgetin blocks transcriptional elongation.

Mass spectrometry analysis combined with in vitro assays reveals that SPINDLY is an O-fucosyltransferase that modifies the growth repressor DELLA and consequently enhances its activity to regulate transcription of target genes.

An automated method for solid-phase polypeptide synthesis capitalizes on rapid amide bond formation to enable the production of multiple traditionally difficult-to-synthesize sequences with both high yield and high purity.

Two screening approaches converge on capzimin, a first-in-class inhibitor of the Rpn11 protease component of the 19S proteasome. Capzimin stabilizes polyubiquitinated substrates, induces the unfolded protein response, and blocks proliferation of cancer cells.

RODEO, an algorithm developed for RiPP natural product discovery, was applied to map out the gene clusters that encode and tailor lasso peptides and led to the identification and characterization of several new lasso peptide topologies.

Integration of heterologous enzymes into the reaction chambers of fungal fatty acid synthases (FASs) demonstrates the capacity of these megaenzymes for engineered production of short- and medium-chain fatty acids and methyl ketones.

Pharmacological chaperones improve folding of destabilized Escherichia coli dihydrofolate reductase (DHFR) and human disease-linked α-galactosidase A (α-GAL) by biasing the kinetic partitioning between folding, aggregation, and degradation. Chaperoning spares DHFR from aggregation and α-GAL from degradation.

In vitro and in silico analysis enables the rational design of fatty acid synthase (FAS)-mediated pathways for the compartmentalized production of desirable fatty acids and a polyketide lactone.

Metabolic labeling of the cell surface with a caged azide sugar enabled cleavage-mediated activation by enzymes overexpressed in cancer cells, allowing enhanced targeted delivery of a doxorubicin conjugate through copper-free click chemistry.

Experimental work and computational modeling together reveal a suite of catalytic roles of the GlcN6P cofactor in the glmS ribozyme, including activation of the nucleophile, electrostatic stabilization, and alignment of the active site.

Design of a proximity-dependent split RNA polymerase system and its optimization by phage-assisted continuous evolution (PACE) enabled the development of a family of activity-dependent split RNA polymerase biosensors regulated by small molecules or light.

In Escherichia coli, replacement of the endogenous tryptophanyl–tRNA synthetase–tRNA pair with its counterpart from Saccharomyces cerevisiae liberates the bacterial counterpart for directed evolution to incorporate unnatural amino acids in both E. coli and eukaryotes.

A gradient of cAMP in developing hippocampal neurons that is important for axon elongation is shaped by spatial differences in phosphodiesterase localization and is maintained by AKAP-anchored PKA, as revealed by using FRET-based biosensors.

Reconstitution experiments using substrates prepared by chemoenzymatic synthesis demonstrate that three LCP family proteins catalyze the ligation of wall teichoic acids to peptidoglycan in the biosynthesis of the Staphylococcus aureus cell wall.

The Ni(ii) affinity of Ni(ii) sensor InrS is attuned to buffered Ni(ii) concentrations, explaining why these two parameters co-vary for different metals over many orders of magnitude.

Spectroscopic studies of allosteric activation of Aurora A kinase using a site-specific infrared probe combined with FRET analysis and molecular dynamics simulations reveals a water-mediated hydrogen bond network in the active site that regulates Aurora A activity.

Discovery and characterization of an unusually permissive C-prenyltransferase provides a biocatalytic route for generating novel prenylated compounds, including daptomycin derivatives with increased potency.

H3K27me3 binding to the EED pocket of the Polycomb repressive complex 2 (PRC2) is required to activate PRC2. An allosteric small-molecule inhibitor of PRC2 was identified that binds to the EED pocket and blocks PRC2 methyltransferase activity in cells.

A pyrrolidine-based small-molecule inhibitor competes with H3K27me3 for binding to EED leading to inactivation of PRC2 and global reduction in H3K27me3 levels.