The development of antibody–drug conjugates remains a challenge in part due to the lack of three-dimensional structural information that must account for the inherent flexibility of antibodies and drug payloads. This Perspective discusses computational methods to guide the design of antibody–drug conjugates.

This Review highlights recent advances in understanding compartmentalized GPCR signaling using chemical biology approaches with a focus on tools for real-time monitoring, site-selective tuning of signaling protein networks and mechanistic screening.

ELOVL6 is an acyltransferase involved in the synthesis of saturated and monounsaturated fatty acids. New work finds that inhibition of ELOVL6 function results in mislocalization of oncogenic KRAS from the plasma membrane, which in turn abolishes KRAS-G12V oncogenesis.

We present a versatile, chemo-ribosomal approach to generating isolable quantities of protein-derived biopolymers containing site-specific backbone modifications, including β-, γ- and δ-peptide linkages. Two of these extended backbones would be difficult to establish by alternative strategies in folded, full-length proteins generated in cells.

Hydrogen exchange–mass spectrometry (HX–MS) is used to qualitatively assess how perturbations such as mutations and ligand binding impact protein ensembles. However, in theory, HX–MS data contain the information necessary to derive residue-level energies of local unfolding (ΔG_op). Now, a method called PIGEON-FEATHER has been developed, which can unambiguously determine residue-level ΔG_op from conventional HX–MS datasets.

Phenotypic screening of an endophyte-derived compound library led to identification of a potent anti-non-small cell lung cancer (NSCLC) lead compound, the 3,4-diisobutyryl derivative of auxarthrol A, uncovering dynein light intermediate chain 1 as a druggable vulnerability and a key regulator of autophagy in NSCLC.

Hydrogen peroxide is a signaling molecule that can also cause damage if its levels are too high. Here, the authors report HyPerFLEX, a fluorescent sensor with tenable colors, to track very low peroxide levels in cellular organelles, even in low-oxygen or highly oxidizing environments.

Duan and Kaushik et al. reveal the structural basis of how Escherichia coli and Thermus thermophilus RNA polymerases initiate transcription from Np₄A alarmones producing Np₄-capped transcripts. The caps form various interactions with a polymerase during initial steps, influencing capping efficiency.

Programming stimulus responsiveness into living systems enables advanced biocomputation. Here, the authors autonomously compile proteins with defined topology that can be site-specifically tethered to and conditionally released from biomaterials and cells following user-specified Boolean logic.

Zhu, Fu, Zhu, Li and colleagues found that the key DNA repair enzyme poly(ADP-ribose) polymerase 1 promotes transcriptional restart after DNA damage through PARylating and stabilizing AFF1 by preventing its ubiquitylation by the E3 ligase Siah1.