Single-molecule imaging prefusion intermediate conformations of MERS-CoV spike trimers in membrane during entry.

Middle East respiratory syndrome coronavirus (MERS-CoV) entry into host cells is mediated by the spike (S) glycoprotein trimer. The S2 domain of spike promotes membrane fusion for MERS entry, but its mechanism of action is currently elusive. Here, we applied real-time single-molecule fluorescence resonance energy transfer (smFRET) imaging to MERS-CoV S virions to identify the prefusion intermediate states of the S2 domain on the pathway to membrane fusion and understand their role in S neutralization by S2 stem-helix-targeted neutralizing antibodies.

Cyclo-stationary distributions of mRNA and Protein counts for random cell division times.

There is a long history of using experimental and computational approaches to study noise in single-cell levels of mRNA and proteins. The noise originates from a myriad of factors: intrinsic processes of gene expression, partitioning errors during division, and extrinsic effects, such as, random cell-cycle times. Although theoretical methods are well developed to analytically understand full statistics of copy numbers for fixed or Erlang distributed cell cycle times, the general problem of random division times is still open.

Catalytically Active Coacervates Sustained Out-of-Equilibrium.

Metabolically active membraneless organelles of extant biology have the capability to maintain their structure under nonequilibrium conditions by leveraging chemical reactions. Herein, we report active coacervates accessed via a mixture of minimal building blocks that featured π-electron rich short peptide, positively charged aldehyde, and a cyclic ketone under nonequilibrium conditions. Peptide bound with the aldehyde by a dynamic covalent bond and demixed to form coacervates through hydrophobic interactions.

Minimal catalytic dissipative assemblies cooperation of an amino acid, a nucleobase precursor and a cofactor.

Functions arising from cooperation between protobiopolymers have fueled the chemical emergence of living matter, which requires a continuous supply of energy to exist in a far-from-equilibrium state. Non-equilibrium conditions imparted by available energy sources have played critical roles in the appearance of complex co-assembled architectures, which exploit the properties of different classes of biopolymers. Such co-assemblies formed from mixtures of nitrogenous heterocycles as protonucleobases and peptide precursors might have acted as early versions of catalytic machinery, capable of sustaining chemical reaction networks.

Optimizing search processes in systems with state toggling: Exact condition delimiting the efficacy of stochastic resetting strategy.

Will the strategy of resetting help a stochastic process to reach its target efficiently, with its environment continually toggling between a strongly favorable and an unfavorable (or weakly favorable) state? A diffusive run-and-tumble motion, transport of molecular motors on or off a filament, and motion under flashing optical traps are special examples of such state toggling. For any general process with toggling under Poisson reset, we derive a mathematical condition for continuous transitions where the advantage rendered by resetting vanishes. For the case of diffusive motion with linear potentials of unequal strength, we present exact solutions, which reveal that there is quite generically a re-entrance of the advantage of resetting as a function of the strength of the strongly favorable potential.

Exact distributions of threshold crossing times of proteins under post-transcriptional regulation by small RNAs.

The timings of several cellular events like cell lysis, cell division, or pore formation in endosomes are regulated by the time taken for the relevant proteins to cross a threshold in number or concentration. Since protein synthesis is stochastic, the threshold crossing time is a first passage problem. The exact distributions of these first passage processes have been obtained recently for unregulated and autoregulated genes.

Pathway-Dependent Catalytic Activity of Short-Peptide-Based Metallozyme: From Promiscuous Activity to Cascade Reaction.

Many natural enzymes contain metal ions as cofactors in the active site for biological activity. However, the pathway of the introduction of metal ions in the earliest protein folds for the emergence of higher catalytic activity remains an intriguing open question. Herein, we demonstrate that pathway-dependent self-assembly of short-peptide-based metallozymes results in differences in catalytic activity.

Feedback driven autonomous cycles of assembly and disassembly from minimal building blocks.

The construction of complex systems by simple chemicals that can display emergent network dynamics might contribute to our understanding of complex behavior from simple organic reactions. Here we design single amino acid/dipeptide-based systems that exhibit multiple periodic changes of (dis)assembly under non-equilibrium conditions in closed system, importantly in the absence of evolved biocatalysts. The two-component based building block exploits pH driven non-covalent assembly and time-delayed accelerated catalysis from self-assembled state to install orthogonal feedback loops with a single batch of reactants.

Elucidating ATP's role as solubilizer of biomolecular aggregate.

Proteins occurring in significantly high concentrations in cellular environments (over 100 mg/ml) and functioning in crowded cytoplasm, often face the prodigious challenges of aggregation which are the pathological hallmark of aging and are critically responsible for a wide spectrum of rising human diseases. Here, we combine a joint-venture of complementary wet-lab experiment and molecular simulation to discern the potential ability of adenosine triphosphate (ATP) as solubilizer of protein aggregates. We show that ATP prevents both condensation of aggregation-prone intrinsically disordered protein Aβ40 and promotes dissolution of preformed aggregates.

Nature of barriers determines first passage times in heterogeneous media.

Intuition suggests that passage times across a region increase with the number of barriers along the path. Can this fail depending on the nature of the barrier? To probe this fundamental question, we exactly solve for the first passage time in general -dimensions for diffusive transport through a spatially patterned array of obstacles - either entropic or energetic, depending on the nature of the obstacles. For energetic barriers, we show that first passage times vary non-monotonically with the number of barriers, while for entropic barriers it increases monotonically.

Non-equilibrium self-assembly for living matter-like properties.

The soft and wet machines of life emerged as the spatially enclosed ensemble of biomolecules with replicating capabilities integrated with metabolic reaction cycles that operate at far-from-equilibrium. A thorough step-by-step synthetic integration of these elements, namely metabolic and replicative properties all confined and operating far-from-equilibrium, can set the stage from which we can ask questions related to the construction of chemical-based evolving systems with living matter-like properties - a monumental endeavour of systems chemistry. The overarching concept of this Review maps the discoveries on this possible integration of reaction networks, self-reproduction and compartmentalization under non-equilibrium conditions.

SARS-CoV-2 spike fusion peptide interaction with phosphatidylserine lipid triggers membrane fusion for viral entry.

Unlabelled: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike is the fusion machine for host cell entry. Still, the mechanism by which spike protein interacts with the target lipid membrane to facilitate membrane fusion during entry is not fully understood. Here, using steady-state membrane fusion and single-molecule fluorescence resonance energy transfer imaging of spike trimers on the surface of SARS-CoV-2 pseudovirion, we directly show that spike protein interacts with phosphatidylserine (PS) lipid in the target membrane for mediating fusion.

Allosteric Control of the Catalytic Properties of Dipeptide-Based Supramolecular Assemblies.

Allostery, as seen in extant biology, governs the activity regulation of enzymes through the redistribution of conformational equilibria upon binding an effector. Herein, a minimal design is demonstrated where a dipeptide can exploit dynamic imine linkage to condense with simple aldehydes to access spherical aggregates as catalytically active states, which facilitates an orthogonal reaction due to the closer proximity of catalytic residues (imidazoles). The allosteric site (amine) of the minimal catalyst can concomitantly bind to an inhibitor via a dynamic exchange, which leads to the alternation of the energy landscape of the self-assembled state, resulting in downregulation of catalytic activity.

Site-selective peptide bond hydrolysis and ligation in water by short peptide-based assemblies.

The evolution of complex chemical inventory from Darwin's nutrient-rich warm pond necessitated rudimentary yet efficient catalytic folds. Short peptides and their self-organized microstructures, ranging from spherical colloids to amyloidogenic aggregates might have played a crucial role in the emergence of contemporary catalytic entities. However, the question of how short peptide fragments had functions akin to contemporary complex enzymes to catalyze cleavage and formation of highly stable peptide bonds that constitute the backbone of all proteins remains an unresolved yet fundamentally important question in terms of the origins of enzymes.

Exact Distribution of the Quantal Content in Synaptic Transmission.

During electrochemical signal transmission through synapses, triggered by an action potential (AP), a stochastic number of synaptic vesicles (SVs), called the "quantal content," release neurotransmitters in the synaptic cleft. It is widely accepted that the quantal content probability distribution is a binomial based on the number of ready-release SVs in the presynaptic terminal. But the latter number itself fluctuates due to its stochastic replenishment, hence the actual distribution of quantal content is unknown.

Estimates of differential toxin expression governing heterogeneous intracellular lifespans of Streptococcus pneumoniae.

Following invasion of the host cell, pore-forming toxins secreted by pathogens compromise vacuole integrity and expose the microbe to diverse intracellular defence mechanisms. However, the quantitative correlation between toxin expression levels and consequent pore dynamics, fostering the intracellular life of pathogens, remains largely unexplored. In this study, using Streptococcus pneumoniae and its secreted pore-forming toxin pneumolysin (Ply) as a model system, we explored various facets of host-pathogen interactions in the host cytosol.

Temporal (Dis)Assembly of Peptide Nanostructures Dictated by Native Multistep Catalytic Transformations.

Emergence of complex catalytic machinery via simple building blocks under non-equilibrium conditions can contribute toward the system level understanding of the extant biocatalytic reaction network that fuels metabolism. Herein, we report temporal (dis)assembly of peptide nanostructures in presence of a cofactor dictated by native multistep cascade transformations. The short peptide can form a dynamic covalent bond with the thermodynamically activated substrate and recruit cofactor hemin to access non-equilibrium catalytic nanostructures (positive feedback).

Exploitation of Catalytic Dyads by Short Peptide-Based Nanotubes for Enantioselective Covalent Catalysis.

Extant enzymes with precisely arranged multiple residues in their three-dimensional binding pockets are capable of exhibiting remarkable stereoselectivity towards a racemic mixture of substrates. However, how early protein folds that possibly featured short peptide fragments facilitated enantioselective catalytic transformations important for the emergence of homochirality still remains an intriguing open question. Herein, enantioselective hydrolysis was shown by short peptide-based nanotubes that could exploit multiple solvent-exposed residues to create chiral binding grooves to covalently interact and subsequently hydrolyse one enantiomer preferentially from a racemic pool.

Temporal Self-Regulation of Mechanical Properties via Catalytic Amyloid Polymers of a Short Peptide.

We report a short peptide that accessed dynamic catalytic polymers to demonstrate four-stage (sol-gel-weak gel-strong gel) temporal self-regulation of its mechanical properties. The peptide exploited its intrinsic catalytic capabilities of manipulating C-C bonds (retro-aldolase-like) that resulted in a nonlinear variation in the catalytic rate. The seven-residue sequence exploited two lysines for binding and cleaving the thermodynamically activated substrate that subsequently led to the self-regulation of the mechanical strengths of the polymerized states as a function of time and reaction progress.

Dual enzyme-powered chemotactic cross β amyloid based functional nanomotors.

Nanomotor chassis constructed from biological precursors and powered by biocatalytic transformations can offer important applications in the future, specifically in emergent biomedical techniques. Herein, cross β amyloid peptide-based nanomotors (amylobots) were prepared from short amyloid peptides. Owing to their remarkable binding capabilities, these soft constructs are able to host dedicated enzymes to catalyze orthogonal substrates for motility and navigation.

Emergence of Photomodulated Protometabolism by Short Peptide-Based Assemblies.

In the early Earth, rudimentary enzymes must have utilized the available light energy source to modulate protometabolic processes. Herein, we report the light-responsive C-C bond manipulation via short peptide-based assemblies bound to the photosensitive molecular cofactor (azo-based photoswitch) where the energy of the light source regulated the binding sites which subsequently modulated the retro-aldolase activity. In the presence of a continual source of high-energy photons, temporal realization of a catalytically more proficient state could be achieved under nonequilibrium conditions.

Emergence of Catalytic Triad by Short Peptide Based Nanofibrillar Assemblies.

Through millions of years of the evolutionary journey, contemporary enzymes observed in extant metabolic pathways have evolved to become specialized, in contrast to their ancestors, which displayed promiscuous activities with wider substrate specificities. However, there remain critical gaps in our understanding of how these early enzymes could show such catalytic versatility despite lacking the complex three-dimensional folds of the existing modern-day enzymes. Herein, we report the emergence of a promiscuous catalytic triad by short amyloid peptide based nanofibers that access paracrystalline folds of β-sheets to expose three residues (lysine, imidazole, and tyrosine) toward solvent.