Biophysics

eLife Assessment

This important study investigates the self-assembly activity of all 109 human death-fold domains. The data collected using advanced microscopy and distributed amphifluoric FRET-based flow cytometry methods are compelling to support the "phase change battery" model that explains how signal amplification can occur without ATP consumption. This paper provides new insight into the thermodynamic control of protein phase behaviors within cells and will be of interest to those studying a variety of biological pathways involved in inflammatory responses and various forms of cell death.

eLife Assessment

This study investigates how RNA molecules modulate phase separation, aggregation, and cytotoxicity of the staphylococcal virulent peptide PSMα3 and the human host‑defence peptide LL‑37 using an array of biophysical and cell‑based assays. If validated, these findings would be important, as they suggest that nucleic acids can tune the material state and bioactivity of amyloids, with implications for host-pathogen interactions and for the design of therapeutics that target phase behaviour. However, the evidence is incomplete: many key claims rest on qualitative imaging and contested assumptions about "functional" amyloids, and the absence of quantitative binding data, phase diagrams, and appropriate controls limits confidence in the conclusions.

eLife Assessment

In their valuable study, Beaudet, Berger and Hendricks provide a mechanistic link between disease-associated tau hyperphosphorylation, loss of cooperative tau envelope formation on microtubules, and dysregulation of axonal transport prior to aggregation. Using complementary in vitro reconstitution and human iPSC-derived neuronal assays with phosphodeficient and phosphomimetic tau constructs targeting 14 disease-relevant sites, the authors convincingly show that phosphorylation state alters tau organization on microtubules and differentially impacts kinesin- and lysosome-based transport. The evidence is solid and well aligned with the conclusions, yet the work could be further strengthened by incorporating additional controls and motor-specific assays to refine the mechanistic depth.

eLife Assessment

This study investigates the folding and unfolding behavior of the doubly knotted protein TrmD-Tm1570, providing insight into the molecular mechanisms underlying protein knotting. The findings reveal multiple unfolding pathways and suggest that the formation of double knots may require chaperone assistance, offering valuable insights into topologically complex proteins. The evidence is convincing, supported by consistent agreement between simulation and experiment, though some aspects of the presentation and experimental scope could be clarified or expanded.

eLife Assessment

This elegant study presents a valuable approach to probing the structural features of the full-length human Hv1 channel as a purified protein, supported by rigorous biochemical assays and spectral FRET analysis, which will interest biophysicists and physiologists studying Hv1 and other ion channels. Overall, the work introduces an interesting labeling strategy and provides methodological observations that are of value in investigating hHV1. However, the analysis appears incomplete, requiring additional structural interpretation and mechanistic insight.

eLife Assessment

This manuscript reports high-resolution cryo-EM structures of a trimethylamine N-oxide demethylase and advances the intriguing hypothesis that the enzyme is bifunctional, coupling TMAO demethylation to formaldehyde capture at a distal tetrahydrofolate-binding site via an enclosed intramolecular tunnel. Supported by biochemical assays and molecular dynamics simulations, the structural findings are valuable and potentially of broad interest, particularly the unusual oligomeric architecture and the proposed conduit for a reactive intermediate. However, the mechanistic framework is considered incomplete, raising substantial concerns regarding the proposed catalytic mechanism, metal/cofactor requirements, and the interpretation of biochemical data supporting formaldehyde channelling.

eLife Assessment

This study addresses a fundamental question in glycobiology by elucidating how a single-site processive enzyme orchestrates the alternating addition of sugars to synthesize complex polysaccharides such as hyaluronan. The findings are compelling, providing a clear mechanistic framework supported by strong experimental validation. Major strengths include the integration of high-resolution structural data with rigorous biochemical analyses, resulting in a well-supported model of hyaluronan assembly.

eLife Assessment

This important study introduces a pulsed laser phase plate that generates stable phase contrast in electron microscopy, offering a practical alternative to continuous-wave designs that suffer from optical instabilities and diffraction artifacts. The experimental results demonstrate a controllable and stable electron phase shift, and the evidence supporting the feasibility of this approach for phase-contrast electron microscopy is convincing. Clarifying the agreement between experiment and theory and further elaborating on possible applications would strengthen the manuscript.

eLife Assessment

This important work examines the effects of side-wall confinement on chemotaxis of swimming bacteria in a shallow microfluidic channel. The authors present convincing experimental evidence, combined with geometric analysis and numerical simulations of simplified models, showing that chemotaxis is enhanced when the distance between the side walls is comparable to the intrinsic radius of chiral circular swimming near open surfaces. This study should be of interest to scientists specializing in bacteria-surface interactions.

eLife Assessment

This study makes a valuable contribution to our understanding of how the lipase regulator ABHD5 may control lipase activity through interactions with lipid droplets and cellular membranes. By combining multiscale molecular dynamics simulations with experimental approaches, the authors provide novel molecular insights into this membrane-protein interaction and present evidence suggesting that the regulatory mechanism depends on protein conformational changes and local membrane remodeling. While much of the evidence supporting the main conclusions is convincing, several aspects of the analysis, interpretation, and discussion remain incomplete. Overall, this work will be of interest to structural and molecular biologists working on lipid metabolism and membrane biophysics.