Biophysics

DEER of Singly Labelled Proteins to Evaluate Supramolecular Packing of Amyloid Fibrils

1. Karen Tsay
2. Asif Equbal
3. Yuanxin Li
4. Tiffany Tsui
5. Songi Han
6. Yann Fichou

• Curated by eLife

  eLife Assessment

  This study presents a valuable methodological contribution exploiting the DEER background decay to quantify supramolecular packing in amyloid fibrils. The evidence is incomplete: the observation of D < 1 is inconsistent with the theoretical lower bound of the model, and it remains unclear whether this reflects a genuine systematic limitation or falls within experimental uncertainty.

Reviewed by eLife

Characterization and modulation of human insulin degrading enzyme conformational dynamics to control enzyme activity

1. Jordan M Mancl
2. Wenguang G Liang
3. Nicholas L Bayhi
4. Hui Wei
5. William Budell
6. Joshua H Mendez
7. Tobin R Sosnick
8. Bridget Carragher
9. Clinton S Potter
10. Wei-Jen Tang

• Curated by eLife

  eLife Assessment

  The manuscript by Mancl et al. provides important mechanistic insights into the conformational dynamics of Insulin Degrading Enzyme (IDE), a zinc metalloprotease involved in the clearance of amyloid peptides. Supported by a compelling combination of time-resolved cryo-EM, SEC-SAXS, enzymatic assays, and both all-atom and coarse-grained simulations, the study reveals an insulin-induced allosteric transition and transient β-sheet interactions underlying IDE's unfoldase activity, thereby refining our understanding of IDE's functional cycle and offering a structural framework for developing substrate-selective modulators of M16 metalloproteases. The latest round of revisions further improves clarity and presentation by updating structural statistics, correcting minor textual inconsistencies, and refining supplemental materials, fully addressing the remaining reviewer comments.

Reviewed by eLife

Structural mechanisms of pump assembly and drug transport in the AcrAB–TolC efflux system

1. Xiaofei Ge
2. Zhiwei Gu
3. Jiawei Wang

• Curated by eLife

  eLife Assessment

  Ge et al here report a structural study of the native tripartite multidrug efflux pump complexes from Escherichia coli that identifies a novel accessory subunit, YbjP, the structure of the native TolC-YbjP-AcrABZ complex, as well as structures of the AcrB protein in L, T, and O conformations. The strength of the structural data is compelling, and the importance of the findings is potentially fundamental. In the revised manuscript, the authors have included additional analysis and made comparisons with pre-existing data which has helped place the data and its impact in the proper context.

Reviewed by eLife

Seeing in the Dark: Intelligent Fourier Light Field Imaging for Bioluminescence Microscopy

1. Luis Felipe Morales Curiel
2. Michael Krieg

Reviewed by PREreview

Unbend, correction of local beam-induced sample motion in cryo-EM images using a 3D spline model

1. Lingli Kong
2. Ximena Zottig
3. Johannes Elferich
4. Nikolaus Grigorieff

• Curated by eLife

  eLife Assessment

  This paper describes Unbend - a new method for measuring and correcting motions in cryo-EM images, with a particular emphasis on more challenging in situ samples such as lamellae and whole cells. The method, which fits a B-spline model using cross-correlation-based local patch alignment of micrograph frames, represents an important tool for the cryo-EM community. The authors elegantly use 2D template matching to provide convincing evidence that Unbend outperforms the previously reported method of Unblur by the same authors. Comparison to alternative programs for motion correction shows smaller gains, but with interesting differences between data sets.

Reviewed by eLife

Lenacapavir-induced Lattice Hyperstabilization is Central to HIV-1 Capsid Failure at the Nuclear Pore Complex and in the Cytoplasm

1. Arpa Hudait
2. Ryan C Burdick
3. Ellie K Bare
4. Vinay K Pathak
5. Gregory A Voth

• Curated by eLife

  eLife Assessment

  This study investigates how the HIV inhibitor lenacapavir influences capsid mechanics and interactions with the nuclear pore complex. It provides important insights into how drug-induced hyperstabilization of the viral shell can compromise its structural integrity during nuclear entry. The modeling is technically sophisticated, and the analyses provide solid support for the mechanistic conclusions.

Reviewed by eLife

Detergent-Triggered Membrane Remodelling Monitored via Intramembrane Fluorescence De-Quenching

1. Claudia M. F. Andrews
2. Christopher M. Hofmair
3. Lauryn Roberts
4. Emily James
5. Katie Morris
6. Kevin Kramm
7. Mark C. Leake
8. Yue Wang
9. Steven D. Quinn

Reviewed by preLights

DNA tensiometer reveals catch-bond detachment kinetics of kinesin-1, -2 and -3

1. Crystal R Noell
2. Tzu-Chen Ma
3. Rui Jiang
4. Scott A McKinley
5. William O Hancock

• Curated by eLife

  eLife Assessment

  The authors use a convincing methodology to investigate the detachment and reattachment kinetics of kinesin-1, 2, and 3 motors against loads oriented parallel to the microtubule. The findings are useful to the field in providing a clearer view of kinesin function under load. However, as the manuscript stands, the conclusions drawn from the experiments, as well as the overall interpretation of the results, are only partially supported by the presented data, and the novelty relative to previous reports appears less clear.

Reviewed by eLife

Dynamic architecture of mycobacterial outer membranes revealed by all-atom simulations

1. Turner P Brown
2. Matthieu Chavent
3. Wonpil Im

• Curated by eLife

  eLife Assessment

  In their study, Brown et. al. provide an important advance in understanding the architecture of the mycobacterial outer membrane. Using all-atom simulations of model mycomembranes, the work reports compelling structural insights into how α-mycolic acids and outer leaflet lipids (PDIM and PAT) shape membrane organisation. The work revealed membrane heterogeneity with ordered inner leaflets and disordered outer leaflets that provide a molecular explanation for the resilience of the mycobacterial envelope.

Reviewed by eLife

Interrogating the structure and function of the human voltage-gated proton channel (hHv1) with a fluorescent noncanonical amino acid

1. Emerson M Carmona
2. William N Zagotta
3. Sharona E Gordon

• Curated by eLife

  eLife Assessment

  This important study presents a convincing methodological approach to probe the structural features of the full-length human Hv1 channel as a purified protein. The method is supported by rigorous biochemical assays and spectral FRET analysis, which will interest biophysicists and physiologists studying Hv1 and other ion channels and membrane proteins. Overall, the work introduces an interesting labeling strategy and provides a methodology that is of value in investigating hHV1 in particular and can be extended to other ion channels. The authors also provide preliminary observations regarding conformational changes induced by zinc.

Reviewed by eLife