Biochemistry

An Automated End-to-End Workflow for Production of Secreted Proteins in Transfected Mammalian Cells

1. Pavee Vasnarungruengkul
2. Michael A. Anaya
3. Annie W. Lam
4. Elisa Gonzalez
5. An Zhang
6. Maxine L. Wang
7. Woj M. Wojtowicz
8. Kai Zinn
9. Jost Vielmetter

Reviewed by Arcadia Science

Kinesin-1 conformational dynamics are controlled by a cargo-sensitive TPR switch

1. Shivam Shukla
2. Jessica A Cross
3. Monika Kish
4. Sathish KN Yadav
5. Johannes F Weijman
6. Laura O’Regan
7. Judith Mantell
8. Ufuk Borucu
9. Xiyue Leng
10. Christiane Schaffitzel
11. Jonathan J Phillips
12. Derek N Woolfson
13. Mark P Dodding

• Curated by eLife

  eLife Assessment

  The revised manuscript by Shukla et al. provides important mechanistic insights into kinesin-1 autoinhibition and cargo-mediated activation. Through a convincing integration of protein engineering, computational modeling, biophysical assays, HDX-MS, and electron microscopy, the study delineates how cargo binding induces an allosteric transition that propagates along the coiled-coil stalk to the motor domains, enhancing MAP7 engagement. The revisions substantially improve clarity, figure annotation, and methodological transparency, leaving the remaining limitations, primarily those inherent to conformational heterogeneity and resolution, appropriately acknowledged. Overall, the updated manuscript presents a coherent mechanism for kinesin-1 activation that will be of broad interest to the motor protein, structural biology, and cell biology communities.

Reviewed by eLife

Elucidating the kinetic and thermodynamic insight into regulation of glycolysis by lactate dehydrogenase and its impact on tricarboxylic acid cycle and oxidative phosphorylation in cancer cells

1. Siying Zeng
2. Yuqi Wang
3. Minfeng Ying
4. Chengmeng Jin
5. Chang Ying
6. Di Wang
7. Hao Wu
8. Xun Hu

• Curated by eLife

  eLife Assessment

  This study presents an assessment of the effect of lactate dehydrogenase (LDH) inhibition on the activity of glycolysis and tricarboxylic acid cycle. The data were collected and analyzed using solid and validated methodology. This paper makes a useful contribution to the field as it considers a control analysis of LDH flux. The findings differ from other published findings likely due to the time course of the incubations used to assess metabolism. While such comparative studies were not presented in the manuscript, the manuscript should be interpreted in light of this critical distinction.

Reviewed by eLife

Interaction hierarchy among Cdv proteins drives recruitment to membrane necks

1. Nicola De Franceschi
2. Alberto Blanch-Jover
3. Cees Dekker

• Curated by eLife

  eLife Assessment

  This important study investigates how the Cdv division proteins of Metallosphaera sedula assemble on and interact with curved membranes in vitro, advancing our understanding of this reduced ESCRT-like machinery. The data provide support for sequential protein recruitment and curvature-dependent enrichment at membrane necks, based on well-controlled reconstitution assays and quantitative analysis. The work establishes a convincing experimental framework for dissecting Cdv-mediated membrane remodeling. The study will be of broad interest to evolutionary and synthetic biologists as well as membrane biophysicists.

Reviewed by eLife

Distinct mechanisms of inhibition of Kv2 potassium channels by tetraethylammonium and RY785

1. Shan Zhang
2. Robyn Stix
3. Esam A Orabi
4. Nathan Bernhardt
5. José D Faraldo-Gómez

• Curated by eLife

  eLife Assessment

  This study represents an important advance in our understanding of how certain inhibitors affect the behavior of voltage gated potassium channels. Robust molecular dynamics simulation and analysis methods lead to a new proposed inhibition mechanism with strength of support being mostly convincing, though computational evidence is limited for some conformations discussed. This study has considerable significance for the fields of ion channel physiology and pharmacology and could aid in development of selective inhibitors for protein targets.

Reviewed by eLife

Mechanism of activation of an ancestral TEC kinase by PIP ₃

1. Eva Krötenheerdt
2. Lucas Piëch
3. Freia von Raußendorf
4. Ronja Reinhardt
5. Nicholas Wedige
6. Hunter G. Nyvall
7. Emma E. Walsh
8. John E. Burke
9. Thomas A. Leonard

Reviewed by Review Commons

Enhancing the protein fitness of interferon-lambda through computational design and glyco-engineering for prophylactic nasal drugs against respiratory viruses

1. Jeongwon Yun
2. Seungju Yang
3. Jae Hyuk Kwon
4. Luiz Felipe Vecchietti
5. Meeyoung Cha
6. Hyun Jung Chung
7. Ji Eun Oh
8. Ho Min Kim

Reviewed by Rapid Reviews Infectious Diseases

Concatenated modular BK channel constructs reveal divergent stoichiometry in gating control by LRRC26 (γ1), pore, and selectivity filter

1. Guanxing Chen
2. Qin Li
3. Kunal Shah
4. Jiusheng Yan

• Curated by eLife

  eLife Assessment

  In this important contribution, Yan and colleagues describe a powerful and compelling strategy to generate concatamers of the BK channel and their fusion constructs with the auxiliary gamma subunits, which allows exploring contributions of individual subunits of the tetrameric channel to its gating and the study of heteromeric channel complexes of defined composition. Distinct examples are presented, which illustrate great diversity in the stoichiometric control of BK channel gating, depending on the site and nature of molecular perturbations. The molecular approaches could be extended to other membrane proteins whose N and C termini face opposite sides of the membrane.

Reviewed by eLife

UV irradiation alters TFAM binding specificity and compaction of DNA

1. Dillon E King
2. Emily E Beard
3. Matthew J Satusky
4. Alex George
5. Ian Ryde
6. Caitlin Johnson
7. Emma L Dolan
8. Yuning Zhang
9. Wei Zhu
10. Hunter Wilkins
11. Evan Corden
12. Susan K Murphy
13. Dorothy A Erie
14. Raluca Gordân
15. Joel Meyer

• Curated by eLife

  eLife Assessment

  Mitochondrial DNA (mtDNA) exhibits a degree of resistance to mutagenesis under genotoxic stress, and this study on the mitochondrial Transcription Factor A (TFAM) presents important data concerning the possible mechanisms involved. The presented data are solid, technically rigorous, and consistent with established literature findings. The experiments are well-executed, providing convincing evidence on the change of TFAM-DNA interactions following UVC irradiation.

Reviewed by eLife

Cul5Wsb2 uses BCL2 proteins as co-receptors to target Bim for degradation

1. Wilhelm Vaysse-Zinkhöfer
2. Enya Marie Catherine Alcindor
3. Nicholas Garaffo
4. David Paul Toczyski

• Curated by eLife

  eLife Assessment

  The study presents a valuable finding on the ubiquitin-dependent regulatory loop in which proapoptotic Bim is targeted to the E3 ubiquitin ligase Cul5-Wsb2-mediated degradation through its sequestration by BCL2 proteins. The conclusions are supported by incomplete evidence and would benefit from additional experiments addressing both the mechanistic understanding and the physiological/cancer-related significance of the study.

Reviewed by eLife