Biochemistry

Purified Zymogens Reveal Mechanisms of Snake Venom Metalloproteinase Auto-Activation

1. Sophie Hall
2. Iara Aimê Cardoso
3. Mark C Wilkinson
4. Maria Molina Carretero
5. Srikanth Lingappa
6. Bronwyn Rand
7. Dakang Shen
8. Johara Boldrini-França
9. Richard Stenner
10. Stefanie K Menzies
11. Georgia Balchin
12. Konrad Kamil Hus
13. Renaud Vincentelli
14. Andrew Mumford
15. Nicholas R Casewell
16. Imre Berger
17. Christiane Schaffitzel

• Curated by eLife

  eLife Assessment

  In this useful paper, the authors present a comprehensive method for the purification of recombinant Snake Venom Metalloproteinases (SVMPs) using the MultiBac expression system, explain the self-activation of the enzymes by Zn2+ incubation, and establish high-throughput screening (HTS) techniques. The authors addressed a key problem: producing a substantial amount of pure and enzymatically active SVMPs required for structural and functional studies. Altogether, this work builds a solid foundation for the large-scale production of active SVMPs for future biochemical and structural characterization as well as for drug discovery, albeit leaving certain caveats about the universal applicability of the described methodology for the production of any recombinant SVMPs.

Reviewed by eLife

Structure of the human CTF18–RFC clamp loader bound to PCNA

1. Giuseppina R Briola
2. Muhammad Tehseen
3. Amani Al-Amodi
4. Grace Young
5. Ammar U Danazumi
6. Phong Quoc Nguyen
7. Christos G Savva
8. Mark Hedglin
9. Samir M Hamdan
10. Alfredo De Biasio

• Curated by eLife

  eLife Assessment

  This paper reports new data on the structure of the human CTF18-RFC clamp loader complex bound to the PCNA clamp. The new and convincing data compliment previous reports of CTF-RFC-PCNA structures and as such, represents an important contribution.

Reviewed by eLife

Common and distinct roles of AMPKγ isoforms in small-molecule activator-stimulated glucose uptake in mouse skeletal muscle

1. Dipsikha Biswas
2. Ever Espino-Gonzalez
3. Danial Ahwazi
4. Jordana B. Freemantle
5. Charline Jomard
6. Jonas Brorson
7. Agnete N. Schou
8. Jean Farup
9. Julien Gondin
10. Jesper Just
11. Marc Foretz
12. Jonas T. Treebak
13. Marianne Agerholm
14. Kei Sakamoto

Reviewed by PREreview

Cryo-EM reveals multiple mechanisms of ribosome inhibition by doxycycline

1. William S. Stuart
2. Michail N. Isupov
3. Mathew McLaren
4. Christopher H. Jenkins
5. Adam Monier
6. Bertram Daum
7. Isobel H. Norville
8. Vicki A.M. Gold
9. Nicholas J. Harmer

Reviewed by preLights

A sulfatide-centered ultra-high resolution magnetic resonance MALDI imaging benchmark dataset for MS1-based lipid annotation tools

1. Lars Gruber
2. Stefan Schmidt
3. Thomas Enzlein
4. Carsten Hopf

Reviewed by GigaScience

Shifting the PPARγ conformational ensemble toward a transcriptionally repressive state improves covalent inhibitor efficacy

1. Liudmyla Arifova
2. Brian S MacTavish
3. Zane Laughlin
4. Mithun Nag Karadi Girdhar
5. Jinsai Shang
6. Min-Hsuan Li
7. Xiaoyu Yu
8. Di Zhu
9. Theodore M Kamenecka
10. Douglas J Kojetin

• Curated by eLife

  eLife Assessment

  This manuscript presents a fundamental advance in our understanding of nuclear receptor pharmacology by expanding on previous work demonstrating dual ligand occupancy in the peroxisome proliferator-activated receptor-gamma (PPARγ). Using a compelling combination of biophysical, biochemical, and cellular approaches, the authors show that covalent inverse agonists with enhanced efficacy shift the receptor conformation toward a transcriptionally repressive state that limits orthosteric ligand co-binding more effectively. This revised manuscript further strengthens support for a proximal, bidirectional allosteric model of dual ligand occupancy by sharpening the distinction between prior and new findings, adding clear conceptual figures, and strengthening statistical rigor.

Reviewed by eLife

RemoteFoldSet: Benchmarking Structural Awareness of Protein Language Models

1. Zinnia Ma
2. Neville P. Bethel

Reviewed by Arcadia Science

A high-resolution analysis of arrestin2 interactions responsible for CCR5 endocytosis

1. Ivana Petrovic
2. Samit Desai
3. Polina Isaikina
4. Layara Akemi Abiko
5. Anne Spang
6. Stephan Grzesiek

• Curated by eLife

  eLife Assessment

  The authors investigate arrestin2-mediated CCR5 endocytosis in the context of clathrin and AP2 contributions. Using an extensive set of NMR experiments, and supported by microscopy and other biophysical assays, the authors provide compelling data on the roles of AP2 and clathrin in CCR5 endocytosis. This important work will appeal to an audience beyond those studying chemokine receptors, including those studying GPCR regulation and trafficking. The distinct role of AP2 and not clathrin will be of particular interest to those studying GPCR internalization mechanisms.

Reviewed by eLife

Mitochondrial Fatty Acid Oxidation is Stimulated by Red Light Irradiation

1. Manuel Alejandro Herrera
2. Camille C. Caldeira da Silva
3. Mauricio S. Baptista
4. Alicia J. Kowaltowski

Reviewed by preLights

Identification of the Regulatory Elements and Protein Substrates of Lysine Acetoacetylation

1. Qianyun Fu
2. Terry Nguyen
3. Bhoj Kumar
4. Parastoo Azadi
5. Y George Zheng

• Curated by eLife

  eLife Assessment

  This useful study reports a method to detect and analyze a novel post-translational modification, lysine acetoacetylation (Kacac), finding it regulates protein metabolism pathways. The study unveils epigenetic modifiers involved in placing this mark, including key histone acetyltransferases such as p300, and concomitant HDACs, which remove the mark. Proteomic and bioinformatics analysis identified many human proteins with Kacac sites, potentially suggesting broad effects on cellular processes and disease mechanisms. The data presented are solid and the study will be of interest to those studying protein and metabolic regulation.

Reviewed by eLife