Molecular Motors Research Articles

Molecular Motors in Myelination and Their Misregulation in Disease.

Molecular motors are cellular components involved in the intracellular transport of organelles and materials to ensure cell homeostasis. This is particularly relevant in neurons, where the synaptic components synthesized in the soma need to travel overlong distances to their destination. They can walk on microtubules (kinesins and dyneins) or actin filaments (myosins), themajor components of cell cytoskeleton. While kinesins mostly perform the anterograde transport of intracellular components toward the plus ends of microtubules located distally in cell processes, cytoplasmic dyneins allow the retrograde flux of intracellular cargo toward the minus ends of microtubules located at the cell soma. Axon myelination represents a major aspect of neuronal maturation and is essential for neuronal function, as it speeds up the transmission of electrical signals. Increasing evidence supports a role for molecular motors in the homeostatic control of myelination. This role includes the trafficking of myelin components along the processes of myelinating cells and local regulation of pathways that ensure axon wrapping. Dysfunctional control of the intracellular transport machinery has thereforebeen linked to several brain pathologies, including demyelinating diseases. These disordersinclude a broad spectrum of conditions characterized by pathological demyelination of axons within the nervous system, ultimately leading to axonal degeneration and neuronal death, withmultiple sclerosis representing the most prevalent and studied condition. This review highlights the involvement of molecular motors in the homeostatic control of myelination. It also discusses studies that have yielded insights into the dysfunctional activity of molecular motors in the pathophysiology of multiple sclerosis.

Bidirectional Molecular Motors by Controlling Threading and Dethreading Pathways of a Linked Rotaxane

AbstractArtificial molecular motors have been presented as models for biological molecular motors. In contrast to the conventional artificial molecular motors that rely on covalent bond rotation, molecular motors with mechanically interlocked molecules (MIMs) have attracted considerable attention owing to their ability to generate significant rotational motion by dynamically shuttling macrocyclic components. The topology of MIM‐type rotational molecular motors is currently limited to catenane structures, which require intricate synthetic procedures that typically produce a low synthetic yield. In this study, we develop a novel class of MIM‐type molecular motors with a rotaxane‐type topology. The switching of the threading/dethreading pathways of the linked rotaxane by protecting/deprotecting the bulky stopper group and changing the solvent polarity enables a net unidirectional rotation of the molecular motor. The threading/dethreading reaction rates were quantitatively evaluated through detailed spectroscopic investigations. Repeated net unidirectional rotation and switching of the direction of rotation were also achieved. Our findings demonstrate that linked rotaxanes can serve as MIM‐type molecular motors with reversible rotational direction controlled by threading/dethreading reactions. These motors hold potential as components of molecular machinery.

MNPs-aptazymes-PtNPs molecular motor biosensor mediated by circular cleavage reactions for the ultrasensitive detection of aflatoxin B1 in real samples

As a significant food safety risk factor linked to cancer, liver diseases, and kidney diseases, aflatoxin B1 (AFB1) necessitates rapid and ultrasensitive detection to mitigate its associated high morbidity and mortality rates. In this study, we developed an advanced biosensor for AFB1 detection by optimizing magnetic nanoparticles (MNPs), platinum nanoparticles (PtNPs), and aptazymes. The resulting MNPs-aptazymes-PtNPs molecular motor biosensor demonstrates promising capabilities for AFB1 detection. Initially, the specific binding between AFB1 and a split aptamer pair facilitates the conversion of intermolecular hybridization of DNAzyme strands into intramolecular hybridization on the MNPs surface. Then the catalytic strands of DNAzyme cleave substrate strands, resulting in PtNPs-containing fragments being released from the MNPs surface and enabling the molecular motor’s movement. The extensive cleavage of substrate strands by the long catalytic strands and the stepwise movement on MNPs surface are driven by the target-specific binding of AFB1. With high peroxidase-mimicking activity, the released PtNPs enable AFB1 detection after short incubation period. The MNPs-aptazymes-PtNPs molecular motor exhibits a sensitivity of 98.95 % and a specificity of 100 %, with a minimum detectable concentration of 300 fg/mL. This thermostabilized, cyclic, and enzyme-free nanomachine demonstrates broad applicability for the detection of various aflatoxins due to its target-empowered, sequence-independent mechanism, possesses excellent guarantee and supervision for enormous Ganoderma products in Shandong province, China.

Unraveling the interplay of kinesin-1, tau, and microtubules in neurodegeneration associated with Alzheimer’s disease

Alzheimer’s disease (AD) is marked by the gradual and age-related deterioration of nerve cells in the central nervous system. The histopathological features observed in the brain affected by AD are the aberrant buildup of extracellular and intracellular amyloid-β and the formation of neurofibrillary tangles consisting of hyperphosphorylated tau protein. Axonal transport is a fundamental process for cargo movement along axons and relies on molecular motors like kinesins and dyneins. Kinesin’s responsibility for transporting crucial cargo within neurons implicates its dysfunction in the impaired axonal transport observed in AD. Impaired axonal transport and dysfunction of molecular motor proteins, along with dysregulated signaling pathways, contribute significantly to synaptic impairment and cognitive decline in AD. Dysregulation in tau, a microtubule-associated protein, emerges as a central player, destabilizing microtubules and disrupting the transport of kinesin-1. Kinesin-1 superfamily members, including kinesin family members 5A, 5B, and 5C, and the kinesin light chain, are intricately linked to AD pathology. However, inconsistencies in the abundance of kinesin family members in AD patients underline the necessity for further exploration into the mechanistic impact of these motor proteins on neurodegeneration and axonal transport disruptions across a spectrum of neurological conditions. This review underscores the significance of kinesin-1’s anterograde transport in AD. It emphasizes the need for investigations into the underlying mechanisms of the impact of motor protein across various neurological conditions. Despite current limitations in scientific literature, our study advocates for targeting kinesin and autophagy dysfunctions as promising avenues for novel therapeutic interventions and diagnostics in AD.

Effect of load-resisting force on photoisomerization mechanism of a single second generation light-driven molecular rotary motor.

A pivotal aspect of molecular motors is their capability to generate load capacity from a single entity. However, few studies have directly characterized the load-resisting force of a single light-driven molecular motor. This research provides a simulation analysis of the load-resisting force for a highly efficient, second-generation molecular motor developed by Feringa et al. We investigate the M-to-P photoinduced nonadiabatic molecular dynamics of 9-(2,3-dihydro-2-methyl-1H-benz[e]inden-1-ylidene)-9H-fluorene utilizing Tully's surface hopping method at the semi-empirical OM2/MRCI level under varying load-resisting forces. The findings indicate that the quantum yield remains relatively stable under forces up to 0.003a.u., with the photoisomerization mechanism functioning typically. Beyond this threshold, the quantum yield declines, and an alternative photoisomerization mechanism emerges, characterized by an inversion of the central double bond's twisting direction. The photoisomerization process stalls when the force attains a critical value of 0.012a.u. Moreover, the average lifetime of the excited state oscillates around that of the unperturbed system. The quantum yield and mean lifetime of the S1 excited state in the absence of external force are recorded at 0.54 and 877.9fs, respectively. In addition, we analyze a time-dependent fluorescence radiation spectrum, confirming the presence of a dark state and significant vibrations, as previously observed experimentally by Conyard et al.

Rab6a enables BICD2/dynein-mediated trafficking of human papillomavirus from the trans-Golgi network during virus entry.

Human papillomaviruses (HPVs) are small, non-enveloped DNA viruses that cause approximately 5% of human cancer. Like most other DNA viruses, HPV traffics to the nucleus during virus entry to successfully infect cells. We show here that HPV utilizes a cellular enzyme, Rab6a, during virus entry to engage the dynein molecular motor for transport along microtubules. Rab6a is required for complex formation between the HPV L2 capsid protein, dynein, and the dynein adaptor BICD2 in the trans-Golgi network (TGN). This complex is required for transport of the incoming virus out of the TGN as it journeys to the nucleus. Our findings identify potential targets for therapeutic approaches.

Tuning the idling molecular motor in muscle to treat nemaline myopathy.

Tuning the idling molecular motor in muscle to treat nemaline myopathy.

The role of kinesin superfamily proteins in hepatocellular carcinoma.

The most prevalent form of primary liver cancer, hepatocellular carcinoma (HCC) poses a significant global health challenge due to its limited therapeutic options. Researchers are currently focused on the complex molecular landscape that governs the initiation and progression of HCC in order to identify new avenues for diagnosis, prognosis, and treatment. In the context of HCC, the Kinesin Superfamily Proteins (KIFs) have become critical regulators of cellular processes, prompting a growing interest in their function among the diverse array of molecular actors implicated in cancer. The KIFs, a family of microtubule-based molecular motors, are renowned for their essential roles in the dynamics of mitotic spindles and intracellular transport. Beyond their well-established functions in normal cellular physiology, emerging evidence indicates that dysregulation of KIFs significantly contributes to the pathogenesis of HCC. Novel therapeutic targets and diagnostic markers are revealed through the unique opportunity to comprehend the complex interplay between KIFs and the molecular events that drive HCC.

Cryo-EM visualizes multiple steps of dynein's activation pathway.

Cytoplasmic dynein-1 (dynein) is an essential molecular motor controlled in part by autoinhibition. We recently identified a structure of partially autoinhibited dynein bound to Lis1, a key dynein regulator mutated in the neurodevelopmental disease lissencephaly. This structure provides an intermediate state in dynein's activation pathway; however, other structural information is needed to fully explain Lis1 function in dynein activation. Here, we used cryo-EM and samples incubated with ATP for different times to reveal novel conformations that we propose represent intermediate states in the dynein's activation pathway. We solved sixteen high-resolution structures, including seven distinct dynein and dynein-Lis1 structures from the same sample. Our data also support a model in which Lis1 relieves dynein autoinhibition by increasing its basal ATP hydrolysis rate and promoting conformations compatible with complex assembly and motility. Together, this analysis advances our understanding of dynein activation and the contribution of Lis1 to this process.

Development of Clinically Viable Non-Muscle Myosin II Small Molecule Inhibitors with Broad Therapeutic Potential.

Research suggests numerous indications, from axon regeneration and cancer, would benefit from a small molecule inhibitor of non-muscle myosin II, a molecular motor that regulates the actin cytoskeleton. Current chemical probe options are very limited and lack sufficient safety for in vivo studies, which we show is primarily due to potent inhibition of cardiac myosin II.Rational design that focused on improving target selectivity over the pan-myosin II inhibitor, blebbistatin, led to the identification of MT-228, a small molecule inhibitor with a wide therapeutic window.High-resolution structure of MT-228 bound to myosin II reveals that selectivity results from a different positioning compared to blebbistatin and an important sequence difference between cardiac and non-muscle myosin II in the inhibitor binding pocket.A single administration of MT-228 shows long-lasting efficacy in animal models of stimulant use disorder, a current unmet and rapidly escalating need with no FDA-approved treatments.

Walking by design: how to build artificial molecular motors made of proteins

Abstract To design an artificial protein-based molecular motor that can autonomously step along a track is a key challenge of protein design and synthetic biology. We lay out a roadmap for how to achieve this aim, based on a modular approach that combines the use of natural, non-motor proteins with de novo design. We define what can be considered to constitute a successful artificial protein motor, identify key steps along the path to achieve these designs, and provide a vision for the future beyond this aim.

Triptycene-Based Tripodal Molecular Platforms.

Molecular platforms are essential components of various surface-mounted molecular devices. Here, we document the synthesis of two universal triptycene-based tripodal pedestals featuring terminal alkynes in the axial position. We showcase their versatility by incorporating them into the structures of diverse functional molecules such as unidirectional light-driven molecular motors, photoswitches, and Brownian molecular rotors using standard cross-coupling reactions. We also present their fundamental physical properties, including acidity constants, data from differential scanning calorimetry, and crystallographic analysis of two parent and five derived structures. Finally, and importantly, we demonstrate that the photochemical properties of selected photoswitch representatives remain uncompromised when fused with tripods.

Impact of dicarboxystilbene impurities on the properties of swift heavy ion latent tracks in PET films

We present a new hypothesis and supporting experimental evidence about the essential role played by small impurities of dicarboxystilbene in the process of UV treatment of PET films irradiated with swift heavy ions (SHI). This treatment both forms highly selective membranes with permeability values comparable to natural ones (the track-UV technique) and sharply accelerates the etching of latent tracks in membrane production (the track-etching technique). We hypothesize (1) that these high permeability value is due to the presence of a molecular motor of an Archimedes screw type in the central part of the latent track formed by photoinduced trans-cis isomerization of dicarboxystilbene molecules that are anchored in helical conformations in the latent track; and (2) that the acceleration of etching rates is due to the preferential accumulation of phenanthrene-type molecules in the central part of the latent track due to the existence of a cyclization channel for photoexcited molecules of cis-dicarboxystilbene. In the presence of alkali solution, phenanthrene molecules release electrons which behave as strong anions in aqueous solution, catalyzing alkaline hydrolysis of PET molecules in the central part of the track. We present experimental observations of photoinduced changes in transmission light intensity after track-UV light treatment of both pristine and SHI irradiated PET films that confirm the presence of trans-cis isomerization of dicarboxystilbene molecules and show their contribution to the formation of new helical conformations in SHI irradiated PET films during the light treatment.

Active Fréedericksz Transition in Active Nematic Droplets

Active nematic liquid crystals have the remarkable ability to spontaneously deform and flow in the absence of any external driving force. While living materials with orientational order, such as the mitotic spindle, can self-assemble in quiescent active phases, reconstituted active systems often display chaotic, periodic, or circulating flows under confinement. Quiescent active nematics are, therefore, quite rare, despite the prediction from active hydrodynamic theory that confinement between two parallel plates can suppress flows. This spontaneous flow transition—named the active Fréedericksz transition by analogy with the conventional Fréedericksz transition in passive nematic liquid crystals under a magnetic field—has been a cornerstone of the field of active matter. Here, we report experimental evidence that confinement in spherical droplets can stabilize the otherwise chaotic dynamics of a 3D extensile active nematics, giving rise to a quiescent—yet still out-of-equilibrium—nematic liquid crystal. The active nematics spontaneously flow when confined in larger droplets. The composite nature of our model system composed of extensile bundles of microtubules and molecular motors dispersed in a passive colloidal liquid crystal allows us to demonstrate how the interplay of activity, nematic elasticity, and confinement impacts the spontaneous flow transition. The critical diameter increases when motor concentration decreases or nematic elasticity increases. Experiments and simulations also demonstrate that the critical confinement depends on the confining geometry, with the critical diameter in droplets being larger than the critical width in channels. Biochemical assays reveal that neither confinement nor nematic elasticity impacts the energy-consumption rate, confirming that the quiescent active phase is the stable out-of-equilibrium phase predicted theoretically. Further experiments in dense arrays of monodisperse droplets show that fluctuations in the droplet composition can smooth the flow transition close to the critical diameter. In conclusion, our work provides experimental validation of the active Fréedericksz transition in 3D active nematics, with potential applications in human health, ecology, and soft robotics. Published by the American Physical Society 2024

Nonthermal fluctuations accelerate biomolecular motors

AbstractIntracellular transport is essential for maintaining cellular function. This process is driven by different mechanisms in prokaryotic and eukaryotic cells. In small prokaryotic cells, diffusion is the primary means of transport, while larger eukaryotic cells also rely on active transport by molecular motors such as kinesin and dynein. Recently, it has become evident that, in addition to diffusion based on thermal fluctuations (Brownian motion), which was conventionally considered a diffusion mechanism within living cells, nonthermal fluctuations generated by metabolic activities play a crucial role in intracellular diffusion. Similarly, while molecular motors have been proposed to exploit thermal fluctuations in the environment following the direct observation and manipulation of single molecules, they have also been reported to utilize nonthermal fluctuations in recent years. This review begins with a brief overview of the historical knowledge of diffusive intracellular transport, which has been extended from the thermal fluctuations to the nonthermal fluctuations generated by metabolic activity. It then introduces recent findings on how nonthermal fluctuations accelerate the motion of molecular motors and discusses future perspectives on the general effects of these fluctuations on molecules in living cells.

The molecular structure of an axle-less F1-ATPase

F1Fo ATP synthase is a molecular rotary motor that can generate ATP using a transmembrane proton motive force. Isolated F1-ATPase catalytic cores can hydrolyse ATP, passing through a series of conformational states involving rotation of the central γ rotor subunit and the opening and closing of the catalytic β subunits. Cooperativity in F1-ATPase has long thought to be conferred through the γ subunit, with three key interaction sites between the γ and β subunits being identified. Single molecule studies have demonstrated that the F1 complexes lacking the γ axle still “rotate” and hydrolyse ATP, but with less efficiency. We solved the cryogenic electron microscopy structure of an axle-less Bacillus sp. PS3 F1-ATPase. The unexpected binding-dwell conformation of the structure in combination with the observed lack of interactions between the axle-less γ and the open β subunit suggests that the complete γ subunit is important for coordinating efficient ATP binding of F1-ATPase.

Kinesin-1 mediates proper ER folding of the CaV1.2 channel and maintains mouse glucose homeostasis.

Glucose-stimulated insulin secretion (GSIS) from pancreatic beta cells is a principal mechanism for systemic glucose homeostasis, of which regulatory mechanisms are still unclear. Here we show that kinesin molecular motor KIF5B is essential for GSIS through maintaining the voltage-gated calcium channel CaV1.2 levels, by facilitating an Hsp70-to-Hsp90 chaperone exchange to pass through the quality control in the endoplasmic reticulum (ER). Phenotypic analyses of KIF5B conditional knockout (cKO) mouse beta cells revealed significant abolishment of glucose-stimulated calcium transients, which altered the behaviors of insulin granules via abnormally stabilized cortical F-actin. KIF5B and Hsp90 colocalize to microdroplets on ER sheets, where CaV1.2 but not Kir6.2 is accumulated. In the absence of KIF5B, CaV1.2 fails to be transferred from Hsp70 to Hsp90 via STIP1, and is likely degraded via the proteasomal pathway. KIF5B and Hsc70 overexpression increased CaV1.2 expression via enhancing its chaperone binding. Thus, ER sheets may serve as the place of KIF5B- and Hsp90-dependent chaperone exchange, which predominantly facilitates CaV1.2 production in beta cells and properly enterprises GSIS against diabetes.

Energetic requirements and mechanistic plasticity in Msp1-mediated substrate extraction from lipid bilayers.

AAA+ proteins are essential molecular motors involved in numerous cellular processes, yet their mechanism of action in extracting membrane proteins from lipid bilayers remains poorly understood. One roadblock for mechanistic studies is the inability to generate subunit specific mutations within these hexameric proteins. Using the mitochondrial AAA+ protein Msp1 as a model, we created covalently linked dimers with varying combinations of wild type and catalytically inactive E193Q mutations. The wide range of ATPase rates in these constructs allows us to probe how Msp1 uses the energy from ATP hydrolysis to perform the thermodynamically unfavorable task of removing a transmembrane helix (TMH) from a lipid bilayer. Our in vitro and in vivo assays reveal a non-linear relationship between ATP hydrolysis and membrane protein extraction, suggesting a minimum ATP hydrolysis rate is required for effective TMH extraction. While structural data often supports a sequential clockwise/2-residue step (SC/2R) mechanism for ATP hydrolysis, our biochemical evidence suggests mechanistic plasticity in how Msp1 coordinates ATP hydrolysis between subunits, potentially allowing for robustness in processing challenging substrates. This study enhances our understanding of how Msp1 coordinates ATP hydrolysis to drive mechanical work and provides foundational insights about the minimum energetic requirements for TMH extraction and the coordination of ATP hydrolysis in AAA+ proteins.

A Multiresponsive Ferrocene-Based Chiral Overcrowded Alkene Twisting Liquid Crystals.

The reversible modulation of chirality has gained significant attention not only for fundamental stereochemical studies but also for numerous applications ranging from liquid crystals (LCs) to molecular motors and machines. This requires the construction of switchable molecules with (multiple) chiral elements in a highly enantioselective manner, which is often a significant synthetic challenge. Here, we show that the dimerization of an easily accessible enantiopure planar chiral ferrocene-indanone building block affords a multi-stimuli-responsive dimer (FcD) with pre-determined double bond geometry, helical chirality, and relative orientation of the two ferrocene motifs in high yield. This intrinsically planar chiral switch can not only undergo thermal or photochemical E/Z isomerization but can also be reversibly and quantitatively oxidized to both a monocationic and a dicationic state which is associated with significant changes in its (chir)optical properties. Specifically, FcD acts as a chiral dopant for cholesteric LCs with a helical twisting power (HTP) of 13 µm-1 which, upon oxidation, drops to near zero, resulting in an unprecedently large redox-tuning of the LC reflection color by up to 84 nm. Due to the straightforward stereoselective synthesis, FcD, and related chiral switches, are envisioned to be powerful building blocks for multi-stimuli-responsive molecular machines and in LC-based materials.

Theoretical insights into rotary mechanism of MotAB in the bacterial flagellar motor

Many bacteria enable locomotion by rotating their flagellum. It has been suggested that this rotation is realized by the rotary motion of the stator unit, MotAB, which is driven by proton transfer across the membrane. Recent cryo-electron microscopy studies have revealed a 5:2 MotAB configuration, in which a MotB dimer is encircled by a ring-shaped MotA pentamer. Although the structure implicates the rotary motion of the MotA wheel around the MotB axle, the molecular mechanisms of rotary motion and how they are coupled with proton transfer across the membrane remain elusive. In this study, we built a structure-based computational model for Campylobacter jejuni MotAB, conducted comprehensive protonation-state-dependent molecular dynamics simulations, and revealed a plausible proton-transfer-coupled rotation pathway. The model assumes rotation-dependent proton transfer, in which proton uptake from the periplasmic side to the conserved aspartic acid in MotB is followed by proton hopping to the MotA proton-carrying site, followed by proton export to the CP. We suggest that, by maintaining two of the proton-carrying sites of MotA in the deprotonated state, the MotA pentamer robustly rotates by ∼36° per proton transfer across the membrane. Our results provide a structure-based mechanistic model of the rotary motion of MotAB in bacterial flagellar motors and provide insights into various ion-driven rotary molecular motors.