Research articles

Condensates have been proposed to create a distinct chemical solvating environment. In vitro condensate screens suggest that condensate chemical environments influence the intracellular distribution of small molecules.

This study reveals β-arrestin-independent GPCR signaling via the cAMP pathway from endosomes, and it delineates a discrete function of β-arrestin in temporally resolving the endosomal signaling phase as a separate peak of cytoplasmic cAMP.

Through directed evolution, the PAM compatibility of the compact Cas9 variant CjCas9 was increased. Evolved CjCas9 shows higher nuclease activity at canonical and non-canonical sites and enables robust in vivo gene editing from single AAV vectors.

Nitric oxide (NO) is a potent vascular signaling agent, but its bioavailability is limited through rapid scavenging reactions. DeMartino et al. characterize the formation and bioactivity of NO-ferroheme, a stable NO analog that forms readily, bypasses scavenging reactions and mediates NO signaling.

Questions remain on the nature of the bioactivity of nitric oxide (NO) synthase signaling despite its wide appreciation. Here the authors describe NO-ferroheme as a vascular signaling species, whose biological activity is unrelated to the release of free nitric oxide, but allows it to travel protected to its main target guanylyl cyclase.

Most miniature Cas12f nucleases have T-rich PAM specificity, restricting their targeting scopes. The cryo-EM structure of the Clostridium novyi Cas12f1 reveals the molecular basis for rare C-rich PAM recognition and enables optimization of sgRNA scaffold to improve CnCas12f1 activity.

Detailed analysis of the structure–activity relationship for cyclin K degraders reveals diverse compounds that acquire glue activity through simultaneous binding to the CDK12 kinase pocket and engagement of several key DDB1 interfacial residues.

Peptide phage display reveals a non-catalytic binding site on the intervening domain of O-GlcNAc transferase. Its roles in substrate recognition, posttranslational modification (PTM) crosstalk and nutrient response provide insight into the function of this cryptic domain.

By solving the cryogenic electron microscopy structures of bacterial calcium-activated potassium channels, Fan et al. report a pathway for blockers to enter the closed pore of the channels through membrane portals rather than through the canonical ion entryway, opening new avenues for drug-targeting this class of channels.

Henneberg et al. developed conformation-specific antibodies enabling probing NEDD8-activated cullin–RING ubiquitin E3 ligase networks in response to extracellular stimuli, metabolic signals and degrader drugs.

Tryptophan hydroxylases have only been known from eukaryotes and are involved in the biosynthesis of serotonin or melatonin. Here, the authors characterize a family of bacterial tryptophan hydroxylases that differ markedly from their eukaryotic counterpart in cofactor and catalytic mechanism.

Homologous to E6AP C-terminus (HECT) E3s forge polyubiquitin chains through multiple reaction steps. A HECT polyubiquitylation cascade was visualized step-by-step, through use of chemical tools and cryo‐EM, and revealed how K48 linkage-specificity is attained by oligomeric UBR5.

Biorefinery of waste biomass provides a sustainable route to produce chemicals and feedstocks. Now, the efficient production of free fatty acids and 3-hydroxypropionic acid from lignocellulose has been achieved via the co-utilization of xylose and glucose from lignocellulose using an engineered yeast.

By developing a genetically encoded biosensor for enzymes that act on O-glycoproteins, Wardman et al. have provided a new method for rapid screening and analysis of these enzymes and performed the first O-glycopeptidase-directed evolution.

Pan, Lu, Feng, Lu et al. demonstrated that CAG repeat expansion RNAs can form gel-like condensates in the cytoplasm and sequester the translation elongation factor eEF2, leading to suppressed global protein synthesis and neurodegeneration-relevant phenotypes.

A fusion and glue platform was developed to determine the cryo-EM structures of GPCRs in diverse states ranging from β2-adrenergic receptors to adhesion receptors.

Functional and length-tunable lipid mimics are computationally designed and co-translationally incorporated into protein in living systems for the gain-of-function study of protein lipidation and the creation of therapeutic candidates.

Most neuropeptides target G-protein-coupled receptors. Now, it has been shown that the tetrapeptide FMRFamide can directly bind and activate a type of ion channel called FMRFamide-activated sodium channels (FaNaCs). This study reports the structure of the FaNaC ion channel in the apo and FMRFamide-bound states and the substrate specificity and possible gating mechanism of FaNaCs.

Cryo-EM structure and dynamics analysis provides a conformational mechanism for tuning of selectivity between calcitonin and amylin receptors through targeted lipid modification of residues 19–22 within the ‘bypass’ motif of amylin peptides.

Asymmetric glucose tracers reveal the simultaneous use and unique benefits of parallel glycolytic pathways. Cells possessing both textbook glycolysis and Entner–Doudoroff glycolysis have a selective advantage in dynamic environments.