Research Briefing |
Featured
-
-
Article
| Open AccessMapping protein dynamics at high spatial resolution with temperature-jump X-ray crystallography
Shifts in temperature alter the structure and dynamics of macromolecules. Now, infra-red laser-induced temperature jump is combined with X-ray crystallography to observe protein structural dynamics in real time. Using this method, motions related to the catalytic cycle of lysozyme, a model enzyme, are visualized at atomic resolution and across broad timescales.
- Alexander M. Wolff
- , Eriko Nango
- & Michael C. Thompson
-
Article
| Open AccessProtein–lipid charge interactions control the folding of outer membrane proteins into asymmetric membranes
Biological membranes are asymmetric bilayers, but little is known about how this asymmetry modulates membrane protein folding or stability. Now, folding and stability assays with bacterial outer membrane proteins reveal an exquisite sensitivity to asymmetric membrane charge distribution and a required matching of protein charge for efficient folding.
- Jonathan M. Machin
- , Antreas C. Kalli
- & Sheena E. Radford
-
Article |
DNAzyme-based faithful probing and pulldown to identify candidate biomarkers of low abundance
The discovery of biomarkers remains challenging owing to a lack of methods sensitive enough to identify such rare molecules. Now, by simultaneously exploiting the catalysis and affinity of a DNAzyme, candidate biomarkers with low abundance in cancers can be pulled down for identification and validation.
- Qinqin Hu
- , Zongxuan Tong
- & Hongzhou Gu
-
Article
| Open AccessAssembling membraneless organelles from de novo designed proteins
Phase separation is being revealed as important in many biological processes. Most attempts to mimic and deconstruct this use engineered natural proteins. Now it is shown that de novo proteins can be designed from first principles to undergo liquid–liquid phase separation in cells, with the potential to organize multi-enzyme pathways.
- Alexander T. Hilditch
- , Andrey Romanyuk
- & Derek N. Woolfson
-
News & Views |
RNA as an off-target for FDA-approved drugs
Medicinal chemistry efforts typically focus on drug–protein interactions and overlook RNA binding as a source of off-target pharmacology. Now, a new method has been developed to map the interactions of small-molecule drugs with RNA in cells and characterize how these interactions can exert functional effects.
- Christopher R. Fullenkamp
- & John S. Schneekloth Jr
-
Article
| Open AccessA SAM analogue-utilizing ribozyme for site-specific RNA alkylation in living cells
Ribozyme-mediated post-transcriptional RNA modification is a powerful method for site-specific RNA labelling and analysis of RNA functions. Now, an alkyltransferase ribozyme—termed SAMURI—has been shown to catalyse the transfer of a propargyl group from a stabilized synthetic S-adenosylmethionine analogue to a specific adenosine on the target RNA within cells.
- Takumi Okuda
- , Ann-Kathrin Lenz
- & Claudia Höbartner
-
Article
| Open AccessPrecisely patterned nanofibres made from extendable protein multiplexes
Molecular systems with coincident cyclic and superhelical symmetry axes have considerable advantages for materials design as they can be lengthened or shortened by changing the length of the monomers. Now a systematic approach to generate modular repeat protein oligomers with combined symmetry that can be extended by repeat propagation has been developed.
- Neville P. Bethel
- , Andrew J. Borst
- & David Baker
-
Article |
Pervasive transcriptome interactions of protein-targeted drugs
Now a reactivity-based RNA profiling strategy can measure the global off-target transcriptome interactions of small-molecule drugs at single-nucleotide resolution. Using this approach, three FDA-approved drugs were evaluated, uncovering pervasive drug–RNA interactions and interactions that perturb RNA functions in cells.
- Linglan Fang
- , Willem A. Velema
- & Eric T. Kool
-
News & Views |
Engineering yeast to produce plant-derived anti-obesity agent
Plants produce a wide range of compounds with important bioactivities. Celastrol, an anti-obesity agent found in the root of certain plants, can now be produced de novo in yeast.
- Jens Nielsen
-
Article
| Open AccessOptical control of ultrafast structural dynamics in a fluorescent protein
Pump–probe measurements conventionally achieve femtosecond time resolution for X-ray crystallography of reactive processes, but the measured structural dynamics are complex. Using coherent control techniques, we show that the ultrafast crystallographic differences of a fluorescent protein are dominated by ground-state vibrational processes that are unconnected to the photoisomerization reaction of the chromophore.
- Christopher D. M. Hutchison
- , James M. Baxter
- & Jasper J. van Thor
-
Article |
Small-molecule autocatalysis drives compartment growth, competition and reproduction
The coupling of autocatalysis to compartment growth and division is a key step in the origin of life. Now it has been shown that compartmentalizing the formose reaction in emulsion droplets leads to several crucial properties of living and evolving systems (growth, division, variation, competition, rudimentary heredity and selection).
- Heng Lu
- , Alex Blokhuis
- & Andrew D. Griffiths
-
News & Views |
Back-to-back cycloadditions in nature
Tandem cycloaddition reactions have significant applications in organic synthetic chemistry. Now, two enzymes are shown to catalyse tandem hetero-Diels–Alder reactions with a synergistic interplay between a calcium cofactor and N-glycan post-translational modifications during the biosynthesis of bistropolone-sesquiterpene secondary metabolites.
- Richiro Ushimaru
- & Ikuro Abe
-
Article
| Open AccessChemical generation of checkpoint inhibitory T cell engagers for the treatment of cancer
Three-protein conjugates, which have so far been produced using protein-engineering strategies, can now be generated using a chemical approach that enables the addition of small-molecule functionality. Checkpoint inhibitory T cell engagers (CiTEs) were assembled and shown to have enhanced in vitro potency compared to a traditional T cell engager.
- Peter A. Szijj
- , Melissa A. Gray
- & Vijay Chudasama
-
Article |
Direct mapping of ligandable tyrosines and lysines in cells with chiral sulfonyl fluoride probes
Most chemoproteomic screening approaches are indirect. Now, a chemoproteomic platform based on chiral sulfonyl fluoride probes has been developed for the direct identification of probe-modified tyrosines and lysines in live cells. Stereoselective modification by structurally diverse probes was observed for 634 tyrosines and lysines across functionally diverse protein sites.
- Ying Chen
- , Gregory B. Craven
- & Jack Taunton
-
Article |
Kinetic control of shape deformations and membrane phase separation inside giant vesicles
The kinetics of liquid–liquid phase separation (LLPS) in cell-like confinements remains poorly understood. Now it has been shown that it involves complex interplay between the incipient phases and the membrane boundary, which arrests phase coarsening, deforms the membrane and couples LLPS with lipid phase separation.
- Wan-Chih Su
- , James C. S. Ho
- & Atul N. Parikh
-
Article
| Open AccessDesign of allosteric sites into rotary motor V1-ATPase by restoring lost function of pseudo-active sites
Allostery produces concerted functions of protein complexes by orchestrating the cooperative work between the constituent subunits. By restoring functions of pseudo-active sites that have been lost through evolution, allosteric sites have now been designed into a rotary molecular motor, V1-ATPase, resulting in its rotation being boosted allosterically.
- Takahiro Kosugi
- , Tatsuya Iida
- & Nobuyasu Koga
-
Article
| Open AccessSite-specific bioorthogonal protein labelling by tetrazine ligation using endogenous β-amino acid dienophiles
An enzymatic reaction installs endogenous β-amino acids in proteins with unique reactivity. Now it has been shows that this reaction can be used for site-specific modification with tetrazine dienophiles to introduce labels onto target proteins. Applications include generation of a radiolabel chelator-modified Her2-binding Affibody and intracellular, fluorescently labelled cell division protein FtsZ.
- Daniel Richter
- , Edgars Lakis
- & Jörn Piel
-
Article |
A DNA-encoded chemical library based on chiral 4-amino-proline enables stereospecific isozyme-selective protein recognition
The design and construction of a stereo-defined DNA-encoded chemical library, featuring the four different 4-amino-proline stereoisomers as a central scaffold, has now enabled the discovery of potent ligands to proteins of pharmaceutical interest. Parallel screening with closely related isoforms (anti-targets) facilitated the isolation of hits with high selectivity ratios.
- Sebastian Oehler
- , Laura Lucaroni
- & Gabriele Bassi
-
News & Views |
Establishing the fundamental rules for genetic code expansion
Genetic code expansion beyond α-amino acids is a major challenge, in which stitching together non-natural building blocks within the ribosome is a critical barrier. Now, the molecular determinants for the efficient incorporation of non-natural amino acids into the ribosome have been unlocked, accelerating ribosomal synthesis.
- Souvik Sinha
- , Mohd Ahsan
- & Giulia Palermo
-
Article |
Tubulin engineering by semi-synthesis reveals that polyglutamylation directs detyrosination
Microtubules carry patterns of post-translational modifications that are important for the regulation of key cellular processes. Now a semi-synthetic method facilitates the production of tubulins with defined post-translational modifications. Using these designer tubulins, polyglutamylation of α-tubulin is found to promote its detyrosination by enhancing the activity of the carboxypeptidase vasohibin/small vasohibin-binding protein.
- Eduard Ebberink
- , Simon Fernandes
- & Charlotte Aumeier
-
Article |
Tandem intermolecular [4 + 2] cycloadditions are catalysed by glycosylated enzymes for natural product biosynthesis
Two glycosylated enzymes, EupfF and PycR1, have now been characterized and shown to independently catalyse the tandem intermolecular [4 + 2] cycloaddition in the biosynthesis of bistropolone-sesquiterpenes. Through analysis of enzyme–substrate co-crystal structures, together with computational and mutational studies, the origins of their catalytic activity and stereoselectivity were elucidated.
- Jiawang Liu
- , Jiayan Lu
- & Youcai Hu
-
Article |
Biosynthesis and biotechnological production of the anti-obesity agent celastrol
Celastrol is a potent anti-obesity agent found in the root of Tripterygium wilfordii, but its medicinal application is compromised by limited availability. Now, by combining plant biochemistry with metabolic engineering and chemistry, the biosynthetic pathway of celastrol has been elucidated and has afforded its scalable production in yeast.
- Yong Zhao
- , Nikolaj L. Hansen
- & Sotirios C. Kampranis
-
Article |
Reversible 2′-OH acylation enhances RNA stability
Stabilization of RNAs for storage, transport and biological application remains a profound challenge. Now, it has been shown that reversible 2′-OH acylation with easily accessible acylimidazoles unlocks efficient protection of RNA. RNA can be deprotected by non-basic nucleophiles or spontaneously in cells to restore RNA functions.
- Linglan Fang
- , Lu Xiao
- & Eric T. Kool
-
Research Briefing |
Synthesis of oligosaccharide libraries for systematic explorations of heparan sulfate sequence space
Challenges in the synthesis of heparan sulfate (HS) glycosaminoglycans have limited access to defined HS oligosaccharides bearing a diverse array of sulfation sequences. A concise, divergent synthetic approach now provides a library of 64 HS tetrasaccharides displaying a comprehensive set of sulfation sequences, offering insight into the elusive sulfation code of glycosaminoglycans.
-
Article |
The dynamics of agonist-β2-adrenergic receptor activation induced by binding of GDP-bound Gs protein
Contrary to agonist binding being the sole driver for β2-adrenergic receptor (β2AR) activation, molecular metadynamics simulations now reveal a distinct activation mechanism. Coupling β2AR with its cognate Gs protein induces considerable structural changes, activating both proteins. Gs opens its GDP binding pocket while β2AR undergoes expansion.
- Amirhossein Mafi
- , Soo-Kyung Kim
- & William A. Goddard III
-
Research Briefing |
Five mutually orthogonal aaRS–tRNA pairs for genetic code expansion
Protein translation is the ultimate paradigm for sequence-defined polymer synthesis. To introduce non-canonical monomers into the genetic code of living organisms, pairs of biomolecules known as aminoacyl-tRNA synthetases (aaRSs) and transfer RNAs (tRNAs) are required. The discovery and engineering of five such pairs, that do not interfere with each other or the aaRS–tRNA pairs of a bacterial host, sets the stage for highly modified genetically encoded biopolymers.
-
Article |
Mass photometric detection and quantification of nanoscale α-synuclein phase separation
The mechanism of α-synuclein amyloid aggregation via liquid–liquid phase separation has so far remained elusive. Now, the existence of nanoscale clusters of α-synuclein in sub-saturated concentrations is observed using mass photometry. These nanoscale clusters can act as precursors to both macroscopic condensate droplets as well as amyloid fibrils.
- Soumik Ray
- , Thomas O. Mason
- & Alexander K. Buell
-
Article |
Quintuply orthogonal pyrrolysyl-tRNA synthetase/tRNAPyl pairs
Mutually orthogonal aminoacyl transfer RNA synthetase/transfer RNA pairs are required for genetically encoding non-canonical amino acids into proteins, as well as for the encoded cellular synthesis of polymers and macrocycles; however, the scalable discovery of such pairs is challenging. A quintuply orthogonal set of pyrrolysyl-tRNA synthetase/pyrrolysyl-tRNA pairs has now been generated through tRNA screening, engineering and directed evolution.
- Adam T. Beattie
- , Daniel L. Dunkelmann
- & Jason W. Chin
-
Article
| Open AccessA locally activatable sensor for robust quantification of organellar glutathione
Fluorescent sensors that are responsive only in a specific subcellular location have remained elusive. Now, a chemogenetic sensing platform has been developed to sense glutathione in a user-defined organelle of interest. These tools enable quantitative studies of subcellular glutathione homeostasis using visible or near-infrared wavelengths.
- Sarah Emmert
- , Gianluca Quargnali
- & Pablo Rivera-Fuentes
-
Article
| Open AccessMetal-dependent enzyme symmetry guides the biosynthetic flux of terpene precursors
The metal-dependent, bifunctional isoprenyl diphosphate synthase PcIDS1 from the leaf beetle Phaedon cochleariae integrates substrate, product and metal-ion concentrations to tune its dynamic reactivity. Now structural and functional analyses reveal that this enzyme uses both catalytic centres to form geranyl pyrophosphate, while one domain is inactivated during farnesyl pyrophosphate production.
- Felix Ecker
- , Abith Vattekkatte
- & Michael Groll
-
Article
| Open AccessAtomistic simulations of the Escherichia coli ribosome provide selection criteria for translationally active substrates
Genetic code expansion to incorporate non-α-amino acid monomers is limited by predictability of monomer reactivities in the context of the ribosome. Now the use of metadynamics simulations of pre-attack monomers in the ribosomal peptidyl transferase centre provides insight on whether an A-site monomer is likely to be reactive.
- Zoe L. Watson
- , Isaac J. Knudson
- & Ara M. Abramyan
-
Article |
Transient water wires mediate selective proton transport in designed channel proteins
Incorporating polar residues into hydrophobic protein channel pores facilitates selective proton transport. Now, classical and multiscale reactive molecular dynamics simulations of designed channels reveal dynamic water wires within the channel lumen that are proton conductive according to structural and functional validation. These results provide some guiding principles for biological and engineered proton conduction.
- Huong T. Kratochvil
- , Laura C. Watkins
- & William F. DeGrado
-
Article |
A high-dimensional microfluidic approach for selection of aptamers with programmable binding affinities
Generating aptamers for use as affinity reagents in analytical applications is important, but SELEX, the standard method for aptamer generation, is unable to select for pre-defined binding affinities. Now, by combining efficient particle display, high-performance microfluidic sorting and high-content bioinformatics, the method ‘Pro-SELEX’ can afford the quantitative generation of aptamers with programmable binding affinities.
- Dingran Chang
- , Zongjie Wang
- & Shana O. Kelley
-
Article
| Open Access2-Oxabicyclo[2.1.1]hexanes as saturated bioisosteres of the ortho-substituted phenyl ring
The ortho-substituted phenyl ring is a basic structural element in chemistry. Now, 2-oxabicyclo[2.1.1]hexanes have been developed as saturated bioisosteres of the ortho-substituted phenyl ring with improved physicochemical properties. Replacement of the phenyl ring with 2-oxabicyclo[2.1.1]hexanes in marketed agrochemicals fluxapyroxad and boscalid improved water solubility, reduced lipophilicity and retained bioactivity.
- Aleksandr Denisenko
- , Pavel Garbuz
- & Pavel K. Mykhailiuk
-
Perspective |
Opportunities and challenges with hyperpolarized bioresponsive probes for functional imaging using magnetic resonance
Bioresponsive hyperpolarized probes contain magnetic resonance signals that can be many orders of magnitude larger than those of common, thermally polarized probes. This Perspective discusses how bioresponsive hyperpolarized probes can be directly linked to biological events to give functional information, enabling the mapping of physiological processes and diseases in real time using magnetic resonance.
- Goran Angelovski
- , Ben J. Tickner
- & Gaoji Wang
-
Article
| Open AccessExpanding the substrate scope of pyrrolysyl-transfer RNA synthetase enzymes to include non-α-amino acids in vitro and in vivo
Ribosomal incorporation of non-α-amino acid monomers into proteins is largely restricted to in vitro translation. Now, pyrrolysyl-transfer RNA synthetase variants have been shown to acylate tRNAs with α-thio acids, malonic acids, and N-formyl amino acids. This work represents a key step towards the programmed ribosomal synthesis of sequence-defined non-protein polymers in cellulo.
- Riley Fricke
- , Cameron V. Swenson
- & Alanna Schepartz
-
News & Views |
Polymeric protagonists for biological processes
Complexity is a hallmark of biological systems, but scientific experiments are typically conducted in simplified conditions. Now, diverse polymers that mimic the local environments of complex biological mixtures have been shown to improve protein folding, stability and function.
- Alana P. Gudinas
- & Danielle J. Mai
-
News & Views |
Illuminating enzyme design using deep learning
From humans designing machines, to machines designing biology, deep learning is turning the tables for assisted exploration of biologically active and diverse protein designs. Now, a deep-learning-based strategy has been used to design artificial enzymes that catalyse a light-emitting reaction.
- Christian Dallago
- & Kevin K. Yang
-
News & Views |
Selecting aptamers with programmed affinities
Constructing aptamers with desired target-binding affinities may lead to new applications but is challenging. Now, a new method using a high-dimensional microfluidic approach enables quantitative isolation of aptamers with programable binding affinities.
- Ping Song
- & Chunhai Fan
-
Article |
Fragmentation and [4 + 3] cycloaddition in sodorifen biosynthesis
The biosynthesis of the methylated sesquiterpene sodorifen, which features a cryptic methylation pattern, has now been studied through extensive labelling experiments and computational chemistry. The methyl group formation is now understood to come from methylene carbons of the substrate farnesyl diphosphate and the absolute configuration of the biosynthetic intermediate presodorifen diphosphate has been revised.
- Houchao Xu
- , Lukas Lauterbach
- & Jeroen S. Dickschat
-
Article
| Open AccessFibril formation and ordering of disordered FUS LC driven by hydrophobic interactions
Protein solutions can undergo liquid–liquid phase separation, by condensing into a dense phase that often resembles liquid droplets, which coexist with a dilute phase. Now it is shown that hydrophobic interactions, specifically at interfaces, can trigger a liquid–solid phase separation of a protein solution.
- Daria Maltseva
- , Sayantan Chatterjee
- & Mischa Bonn
-
Article
| Open AccessIn vitro selection of macrocyclic peptide inhibitors containing cyclic γ2,4-amino acids targeting the SARS-CoV-2 main protease
In vitro screening of a ribosomally synthesized macrocyclic peptide library containing cyclic γ2,4-amino acids (cγAA) afforded the discovery of potent inhibitors of the SARS-CoV-2 main protease (Mpro). A co-crystal structure revealed the contribution of this cγAA to Mpro binding and the proteolytic stability of these macrocycles.
- Takashi Miura
- , Tika R. Malla
- & Hiroaki Suga
-
Article
| Open AccessIn vivo metallophilic self-assembly of a light-activated anticancer drug
The metallophilic interaction between cyclometalated palladium complexes can facilitate supramolecular nanostructure formation in living mice, providing a phototoxic prodrug with a long circulation time and high tumour-targeting efficiency. Upon green light irradiation, this palladium-based drug destroys solid tumours, leaving non-irradiated organs intact.
- Xue-Quan Zhou
- , Peiyuan Wang
- & Sylvestre Bonnet
-
In Your Element |
Looking into luciferin
Organisms that glow are perhaps eerie. Vadim Viviani ponders on the luciferin–luciferase systems responsible for their intriguing bioluminescence.
- Vadim R. Viviani
-
Article |
Spatiotemporal functional assembly of split protein pairs through a light-activated SpyLigation
Techniques to specifically modulate protein activity are needed to interrogate spatial effects in cellular processes. A genetically encoded method for site-specific protein–protein conjugation based on a photoclick chemical reaction has now been developed. This method permits rapid and irreversible reassembly of bioactive proteins from non-functional split fragment pairs with full spatiotemporal control in solution, biomaterials and living mammalian cells.
- Emily R. Ruskowitz
- , Brizzia G. Munoz-Robles
- & Cole A. DeForest
-
In Your Element |
Giving the green light
Jane Liao and Allie C. Obermeyer explore the discovery, modification and applications of green fluorescent protein, best known for its use as a tool to cast light on cellular processes.
- Jane Liao
- & Allie C. Obermeyer
-
Article |
OregonFluor enables quantitative intracellular paired agent imaging to assess drug target availability in live cells and tissues
Water-soluble, cell-permeable, inert fluorescent tags called OregonFluors have been developed to withstand environmental changes while resistant towards non-specific binding with subcellular structures. These tags enable quantitative imaging of drug target availability in cells and tissues, providing a route for the future assessment of personalized therapies.
- Lei G. Wang
- , Antonio R. Montaño
- & Summer L. Gibbs
-
News & Views |
Illuminating anti-ageing
Therapies that destroy senescent cells could be used to alleviate age-related disease, yet conventional drugs often suffer from low selectivity and unwanted side effects. Now, a photosensitive agent has been developed that is activated in situ in senescent cells, enabling their selective elimination.
- Yunjie Xu
- , Jong Seung Kim
- & Mingle Li
-
Article |
Connecting the geometric and electronic structures of the nitrogenase iron–molybdenum cofactor through site-selective 57Fe labelling
The molybdenum nitrogenase catalytic cofactor is composed of seven high-spin Fe sites making it difficult to study spectroscopically. Now it has been shown that 57Fe can be incorporated into a single site and that such site-selectively labelled samples provide insights into the cofactor’s electronic structure and the mechanism of biological nitrogen fixation.
- Edward D. Badding
- , Suppachai Srisantitham
- & Daniel L. M. Suess
Browse narrower subjects
- Biochemistry
- Biological techniques
- Biophysics
- Biotechnology
- Cancer
- Cell biology
- Chemical biology
- Computational biology and bioinformatics
- Developmental biology
- Drug discovery
- Ecology
- Evolution
- Genetics
- Immunology
- Microbiology
- Molecular biology
- Neuroscience
- Physiology
- Plant sciences
- Psychology
- Stem cells
- Structural biology
- Systems biology
- Zoology