Article
Chemistry, Multidisciplinary
Chong Zuo, Ruichao Ding, Xiangwei Wu, Yuanxia Wang, Guo-Chao Chu, Lu-Jun Liang, Huasong Ai, Ze-Bin Tong, Junxiong Mao, Qingyun Zheng, Tian Wang, Zichen Li, Lei Liu, Demeng Sun
Summary: This study reports a new method for protein labeling and semi-synthesis using SrtA-mediated ligation. The method shows broad tolerance for LPxT motifs and enables efficient and irreversible connection of protein substrates.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Review
Biochemistry & Molecular Biology
Stephen G. Sligar, Ilia G. Denisov
Summary: Membrane proteins play crucial roles in biological processes, but are often challenging to study. The NanoDisc platform offers a self-assembled system that solubilizes membrane proteins in water while maintaining their functional activity. NanoDiscs have utility in studying cellular signaling complexes and delivering drugs, showcasing their versatility in biochemistry and pharmacology.
Article
Chemistry, Multidisciplinary
Ilkoo Noh, Zhongyuan Guo, Jiarong Zhou, Weiwei Gao, Ronnie H. Fang, Liangfang Zhang
Summary: Vaccination with nanovaccines based on bacterial outer membrane has shown promising results in protecting against antibiotic-resistant infections. The cellular nanodiscs developed in this study effectively interact with antigen-presenting cells and can efficiently transport to lymph nodes. In a murine model, the nanovaccine elicits potent immune responses and confers protection against subsequent lung infection.
Article
Biochemistry & Molecular Biology
Laura E. Sperl, Florian Ruehrnoessl, Anita Schiller, Martin Haslbeck, Franz Hagn
Summary: The study used NMR and HDX-MS techniques in lipid nanodiscs to investigate the high-resolution structural changes of Bak in a membrane environment, providing important insights into Bak-mediated membrane permeabilization. Experimental results showed that the alpha-helix 1 element in Bak is a metastable structure contributing to its pro-apoptotic features.
Article
Chemistry, Physical
Ramsia Geisler, Martin Cramer Pedersen, Natalie Preisig, Yvonne Hannappel, Sylvain Prevost, Rajeev Dattani, Lise Arleth, Thomas Hellweg
Summary: The study investigates the self-assembly and morphological transition of nm-sized discoidal lipid nanoparticles composed of beta-aescin and the phospholipid DMPC. The transition of the lipid nanoparticles is mainly triggered by the phase state change of the phospholipid, resulting in different final morphologies depending on the phospholipid-to-saponin ratio and temperature. Techniques like SAXS, TEM, and FFEM are used to analyze the nano-particles at different length scales, with models applied to evaluate possible geometries and molecular mixing.
Article
Developmental Biology
Miquel Sendra, Juan de Dios Hourcade, Susana Temino, Antonio J. Sarabia, Oscar H. Ocana, Miguel Torres, Jorge N. Dominguez
Summary: Tracing and manipulating cells in embryos are essential to understand development. Lipophilic dye microinjections, viral transfection, and iontophoresis have been key techniques for mapping the origin of progenitor cells. Here, the researchers used cell-permeant Cre recombinase microinjection to perform lineage tracing and gene ablation. The tool will help identify cell progenitors and gene networks involved in organ development and understand the origin of congenital defects.
Article
Chemistry, Analytical
Jamal Ahmed Cheema, Nihan Aydemir, Colm Carraher, Roshan Khadka, Damon Colbert, Harris T. Lin, Andrew Nelson, Andrew Kralicek, Jadranka Travas-Sejdic
Summary: This study presents an electrochemical sensing methodology using an insect odorant receptor (iOR) as the biological recognition element for detection of odorant compounds. The fruit fly Drosophila melanogaster's Or22a was used as a model receptor and shown to be functionally active towards its known ligand (ethyl hexanoate). The EIS sensor exhibited high sensitivity and specificity towards ethyl hexanoate with a detection limit of 5.5 fM, with neutron reflectometry studies providing evidence of conformational changes in the receptor upon ligand binding.
SENSORS AND ACTUATORS B-CHEMICAL
(2021)
Article
Materials Science, Multidisciplinary
Milena Timcenko, Anton A. A. Autzen, Henriette E. Autzen
Summary: Amphiphilic copolymers have the potential to extract membrane proteins directly from lipid bilayers into native nanodiscs. However, many such copolymers are polyanionic and sensitive to divalent cations. In this study, we characterized the Ca2+ and Mg2+ sensitivity of poly(acrylic acid-co-styrene) (AASTY) copolymers and found that divalent cations promote aggregation and precipitation of both free and lipid-bound copolymers. Excess, free copolymer acts as a cation sink that protects nanodiscs from Ca2+-induced aggregation. Removal of the free copolymer induces aggregation, which can be mitigated by KCl. Our findings also revealed that the size of the nanodiscs is dynamic and dependent on lipid concentration.
ACS APPLIED POLYMER MATERIALS
(2022)
Article
Chemistry, Medicinal
Feng Jiang, Chengteng Cai, Xiumin Wang, Shoufa Han
Summary: Signal-on labeling and photodynamic destruction of Staphylococcus aureus (S. aureus) were achieved using self-quenched optical probes and Sortase A-mediated cleavage of a substrate motif. This dual biomarked-enabled peptidoglycan labeling approach shows potential for targeted therapy and diagnosis of SrtA-positive bacteria associated with various diseases.
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS
(2023)
Article
Chemistry, Multidisciplinary
Qingyun Yang, Yingxiao Gao, Xia Liu, Yihang Xiao, Mingxuan Wu
Summary: A new method for editing the N-tail of histone H3 using sortase mediated metathesis (SMM) is reported. This method expands the application scope of sortase and offers a strategy for precise editing of PTMs of cellular histone H3.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Chemistry, Multidisciplinary
Xinying Jia, Yanni K. -Y. Chin, Alan H. H. Zhang, Theo Crawford, Yifei Zhu, Nicholas L. L. Fletcher, Zihan Zhou, Brett R. R. Hamilton, Martin Stroet, Kristofer J. J. Thurecht, Mehdi Mobli
Summary: The authors report the engineering of self-cyclizing 'autocyclase' proteins to generate macrocyclic peptides and proteins with favorable reaction kinetics for suppressing polymerization. Macrocyclisation of proteins and peptides results in a remarkable increase in structural stability, making cyclic peptides and proteins of great interest in drug discovery. The engineering of a self-cyclising autocyclase protein provides a simple, alternative way to access a vast diversity of macrocyclic biomolecules.
COMMUNICATIONS CHEMISTRY
(2023)
Article
Chemistry, Multidisciplinary
Mengmeng Zheng, Fa-Jie Chen, Kaicheng Li, Rahi M. Reja, Fredrik Haeffner, Jianmin Gao
Summary: Binding via reversible covalent bond formation is a powerful mechanism to enhance the potency of synthetic inhibitors. This study reports the development of chemically modified phage libraries that incorporate 2-acetylphenylboronic acid (APBA) for reversible binding, and demonstrates their utility against two protein targets, showing high potency and specificity.
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(2022)
Article
Biophysics
Tongtong Tian, Jinzhi Zhao, Yuning Wang, Binxiao Li, Liang Qiao, Kun Zhang, Baohong Liu
Summary: The research developed a novel digital single-particle imaging method to quantify sortase A (SrtA) activity by assembling and counting gold nanoparticles, providing a facile and novel analytical tool for understanding infection mechanisms and pharmaceutical development.
BIOSENSORS & BIOELECTRONICS
(2021)
Article
Developmental Biology
Sara Caviglia, Iris A. Unterweger, Akvile Gasiunaite, Alexandre E. Vanoosthuyse, Francesco Cutrale, Le A. Trinh, Scott E. Fraser, Stephan C. F. Neuhauss, Elke A. Ober
Summary: Visualizing cell shapes and interactions is crucial for understanding organ development and repair. However, integrating multi-fluorophore information into developing zebrafish tissues is challenging. In this study, the researchers developed FRaeppli, a method for efficient cell shape analysis and tracking using specific fluorescent proteins.
Article
Chemistry, Multidisciplinary
Vishal Maingi, Paul W. K. Rothemund
Summary: The study found that charge neutralization of DNA scaffolds was more important than the total hydrophobic content of their modifications in achieving bulk bilayer behavior. Additionally, complete charge neutralization of DNA scaffolds enabled better lipid binding and more stable bilayers. In conclusion, the simulations provide a guide for the design of DNA-scaffolded nanodiscs suitable for studying membrane proteins.
Article
Chemistry, Analytical
Pattanapong Thangsunan, Nidhi Lal, Christian Tiede, Sophie Moul, James Robinson, Margaret A. Knowles, Peter G. Stockley, Paul A. Beales, Darren C. Tomlinson, Michael J. McPherson, Paul A. Millner
Summary: FGFR3 is a key receptor protein with potential biomarker role in bladder cancer. Affimer reagents targeting FGFR3 showed strong binding affinity and were used for fabrication of impedimetric electrochemical biosensors. These biosensors were capable of detecting FGFR3 protein at sub-pM to nM concentrations in different solutions.
SENSORS AND ACTUATORS B-CHEMICAL
(2021)
Article
Chemistry, Physical
Ylenia Miele, Stephen J. Jones, Federico Rossi, Paul A. Beales, Annette F. Taylor
Summary: The transmission of chemical signals through extracellular solution is important in cellular biological systems and lipid vesicles applications. This study investigates chemical communication in synthetic micro-and nanovesicles containing urease in a solution of urea and acid. Through experiments and simulations, it is demonstrated that the fast transport of ammonia to the external solution controls the pH-time profile and synchronizes the timing of the pH clock reaction in a heterogeneous population of vesicles. This research shows how the rate of production and emission of a small basic product controls pH changes in active vesicles with different sizes and enzyme amounts, which may have applications in bioreactors or healthcare.
JOURNAL OF PHYSICAL CHEMISTRY LETTERS
(2022)
Article
Chemistry, Physical
Rashmi Seneviratne, Rosa Catania, Michael Rappolt, Lars J. C. Jeuken, Paul A. Beales
Summary: Lipids and block copolymers can self-assemble into vesicles, and combining them allows for the optimization of vesicle membranes for bionanotechnology applications. Increasing the polymer fraction in the mixture improves membrane mixing but decreases membrane fluidity in hybrid vesicles.
Review
Materials Science, Biomaterials
Matthew P. Culbert, James P. Warren, Andrew R. Dixon, Hazel L. Fermor, Paul A. Beales, Ruth K. Wilcox
Summary: Back pain can affect health and mobility, and nucleus augmentation aims to restore the intervertebral disc to its healthy state through minimally invasive methods. Injectable hydrogels are commonly used materials for nucleus augmentation, and the mechanical properties and biological compatibility of these materials are important factors to consider in their development.
BIOMATERIALS SCIENCE
(2022)
Article
Chemistry, Multidisciplinary
Matthew J. Tamasi, Roshan A. Patel, Carlos H. Borca, Shashank Kosuri, Heloise Mugnier, Rahul Upadhya, N. Sanjeeva Murthy, Michael A. Webb, Adam J. Gormley
Summary: This study reports a strategy for designing protein-stabilizing copolymers based on active machine learning, which successfully identified copolymers that preserve or even enhance the activity of three chemically distinct enzymes under thermal denaturing conditions. Although systematic screening results were mixed, active learning appropriately identified unique and effective copolymer chemistries for the stabilization of each enzyme, broadening the capabilities to design fit-for-purpose synthetic copolymers.
ADVANCED MATERIALS
(2022)
Article
Polymer Science
Rosa Catania, Jonathan Machin, Michael Rappolt, Stephen P. Muench, Paul A. Beales, Lars J. C. Jeuken
Summary: In this study, we demonstrate a new approach of extracting membrane proteins using styrene-maleic acid (SMA) that allows for direct transfer of cytochrome bo(3) into hybrid vesicles (HVs). This method is efficient, avoiding the need for detergent, long incubation times, or mechanical disruption. HVs are shown to be more suitable than liposomes for membrane protein incorporation from SMA lipid particles. Furthermore, this transfer method is not limited to a single membrane protein and can be used with complex membrane protein mixtures.
Article
Biophysics
Marcos Arribas Perez, Paul A. Beales
Summary: Membrane fusion is a way to increase the complexity of model membrane systems. In this study, silica nanoparticles were used to fuse liquid-disordered DOPC giant unilamellar vesicles (GUVs) and liquid-ordered DPPC:cholesterol (7:3) GUVs. After fusion, GUVs displayed large membrane domains, indicating lipid redistribution between phase domains. Real-time confocal microscopy revealed the dynamics of asymmetric membrane domain formation in hemifused GUVs and the modulation of the inner membrane leaflet phase state by the DOPC-rich liquid-disordered domains in the outer monolayer.
BIOPHYSICAL JOURNAL
(2023)
Article
Chemistry, Multidisciplinary
Jonathan P. Dolan, Darren C. Machin, Simone Dedola, Robert A. Field, Michael E. Webb, W. Bruce Turnbull
Summary: This study describes the chemoenzymatic synthesis of dual N- and C-terminal-functionalized cholera toxin B subunit (CTB) glycoconjugates. By using sortase-mediated ligation, MUC1(Tn) peptides are conjugated to the C-terminus of CTB, allowing for high-density display of the MUC1(Tn) epitopes.
FRONTIERS IN CHEMISTRY
(2022)
Article
Biochemical Research Methods
Holly E. Morgan, Zoe L. P. Arnott, Tomasz P. Kaminski, W. Bruce Turnbull, Michael E. Webb
Summary: This paper presents a method using evolved sortase variants and depsipeptide substrates for site-specific labeling, which expands the biochemical protein labeling toolkit. An orthogonal pair of enzymes accepting LPEToG and LPESoG depsipeptides has been identified and successfully applied to dual N-terminal labeling.
BIOCONJUGATE CHEMISTRY
(2022)
Article
Chemistry, Multidisciplinary
Rashmi Seneviratne, Georgina Coates, Zexi Xu, Caitlin E. Cornell, Rebecca F. Thompson, Amin Sadeghpour, Daniel P. Maskell, Lars J. C. Jeuken, Michael Rappolt, Paul A. Beales
Summary: In this study, the detailed structural information about hybrid vesicles composed of phospholipids and block-copolymers was obtained using small angle X-ray scattering and cryo-electron tomography. The findings reveal that hybrid vesicles with different ratios of lipids and polymers have different membrane thicknesses, indicating the homogeneous mixing of lipids and polymers. This research is important for understanding the structural properties of hybrid membranes.
Article
Polymer Science
James P. Warren, Matthew P. Culbert, Danielle E. Miles, Steven Maude, Ruth K. Wilcox, Paul A. Beales
Summary: Self-assembling peptides are a versatile biomaterial that can form self-supporting hydrogels. Balancing attractive and repulsive forces through peptide charge and hydrogen bonding is crucial for successful hydrogel formation. A net peptide charge of +/-2 and modulating hydrogen bonding can optimize the assembly and viscoelastic properties of the hydrogel. These findings demonstrate the potential of controlling self-assembly mechanisms to create structures with desired properties.
Article
Chemistry, Multidisciplinary
Ryan McBerney, Jonathan P. Dolan, Emma E. Cawood, Michael E. Webb, W. Bruce Turnbull
Summary: This study presents a site-specific bioorthogonal approach for synthesizing homogeneous glycoproteins. By developing a bifunctional bioorthogonal linker and functionalizing reducing sugars and glycan derivatives, synthetic neoglycoproteins were successfully produced as inhibitors of cholera toxin, demonstrating the potential of this method for creating homogeneous glycoproteins.
Article
Chemistry, Organic
Asif Fazal, Jake Wheeler, Michael E. Webb, Ryan F. Seipke
Summary: SurE is a peptide cyclase essential for the production of surugamide antibiotics. However, the substrate specificity of this enzyme is not well understood. In order to address this issue, researchers developed a new assay to investigate the chemical space of the N-terminal substrate residue.
ORGANIC & BIOMOLECULAR CHEMISTRY
(2022)
Article
Chemistry, Physical
Marcos Arribas Perez, Paul A. Beales
Summary: The formation of a protein corona significantly alters the behavior of nanoparticles, including increasing their size and changing their morphological response. Additionally, the protein corona weakens the effect of nanoparticles on lipid membranes.
Review
Chemistry, Multidisciplinary
Holly E. Morgan, W. Bruce Turnbull, Michael E. Webb
Summary: Site-specific protein modification is a widely-used biochemical tool, but there are challenges in its development. This review focuses on the engineering of peptide ligases and their substrates to address these challenges, and explores techniques to improve reaction efficiency and expand substrate scope.
CHEMICAL SOCIETY REVIEWS
(2022)
