Targeted Optimization of a Protein Nanomachine for Operation in Biohybrid Devices

Amrute-Nayak, Mamta; Diensthuber, Ralph P.; Walter, Steffen; Kathmann, Daniela; Hartmann, Falk K.; Fedorov, Roman; Urbanke, Claus; Manstein, Dietmar J.; Brenner, Bernhard; Tsiavaliaris, Georgios

doi:10.1002/anie.200905200

Cited by 19 publications

(18 citation statements)

References 37 publications

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“…The functionalization of biohybrid devices that exploit actomyosin-based cargo transport for molecular diagnostics and other nanotechnological applications (Amrute-Nayak et al, 2010; Lard et al, 2013; Persson et al, 2013) is another area that will certainly benefit from the use of EMD 57033 and the development of compounds that stabilize their target proteins against stress-induced denaturation, precipitation, and proteolytic degradation. The effective and simple long-term storage of chip-based nanobiosensors, where actomyosin-driven transport substitutes microfluidics and forms the basis for novel detection schemes, is a precondition for the commercial viability of such devices.…”

Section: Discussionmentioning

confidence: 99%

Small molecule-mediated refolding and activation of myosin motor function

Radke

Taft

Stapel

et al. 2014

eLife

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The small molecule EMD 57033 has been shown to stimulate the actomyosin ATPase activity and contractility of myofilaments. Here, we show that EMD 57033 binds to an allosteric pocket in the myosin motor domain. EMD 57033-binding protects myosin against heat stress and thermal denaturation. In the presence of EMD 57033, ATP hydrolysis, coupling between actin and nucleotide binding sites, and actin affinity in the presence of ATP are increased more than 10-fold. Addition of EMD 57033 to heat-inactivated β-cardiac myosin is followed by refolding and reactivation of ATPase and motile activities. In heat-stressed cardiomyocytes expression of the stress-marker atrial natriuretic peptide is suppressed by EMD 57033. Thus, EMD 57033 displays a much wider spectrum of activities than those previously associated with small, drug-like compounds. Allosteric effectors that mediate refolding and enhance enzymatic function have the potential to improve the treatment of heart failure, myopathies, and protein misfolding diseases.DOI: http://dx.doi.org/10.7554/eLife.01603.001

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Section: Discussionmentioning

confidence: 99%

Small molecule-mediated refolding and activation of myosin motor function

Radke

Taft

Stapel

et al. 2014

eLife

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“…In this context, investigators have regularly relied on Diatrack’s dedicated functionalities for in vitro motility studies (‘Particle production…’ ‘in vitro motility assay’). For example, Taft et al demonstrated that myosin-5b is a processive motor 55 and Amrute-Nayak et al succeeded in engineering single and double headed myosin-5 motors, whose mechanochemical properties (force, speed, processivity) could be systematically controlled by variations in ATP, ionic strength, and Mg 2+ -ion concentrations 56 . Further, the effects of small molecules on myosins can be easily assessed in the in vitro motility assay by characterizing the changes in the velocity of filaments.…”

Section: Survey Of Applicationsmentioning

confidence: 99%

Diatrack particle tracking software: Review of applications and performance evaluation

Vallotton

Oijen

Whitchurch

et al. 2017

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Object tracking is an instrumental tool supporting studies of cellular trafficking. There are three challenges in object tracking: the identification of targets; the precise determination of their position and boundaries, and the assembly of correct trajectories. This last challenge is particularly relevant when dealing with densely populated images with low signal-to-noise ratios - conditions that are often encountered in applications such as organelle tracking, virus particle tracking, or single-molecule imaging. We have developed a set of methods that can handle a wide variety of signal complexities. They are compiled into a free software package called Diatrack. Here we review its main features and utility in a range of applications, providing a survey of the dynamic imaging field together with recommendations for effective use. The performance of our framework is shown to compare favorably to a wide selection of custom-developed algorithms, whether in terms of localization precision, processing speed, or correctness of tracks.

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“…For further development of point-of-care diagnostic devices, it will be important to perform in depth studies of the mechanisms that limit the capability of engineered cytoskeletal filament shuttles to transport various cargoes, from single macromolecules to cells. Other important developments would be new approaches for optimizing (Amrute-Nayak et al 2010) and stabilizing the protein components in order to increase their shelf-life and methods to ensure the compatibility of motor function with complex fluid environments or alternatively, bypassing this complication by innovative approaches. Additionally, it will be important to select appropriate applications for the different motor system to optimally utilize the high cargo-carrying capacity of microtubule kinesin and the high speed and increased capability of miniaturization associated with myosin II propelled actin filaments.…”

Section: Developments From 2005 and Onwardsmentioning

confidence: 99%

Translational actomyosin research: fundamental insights and applications hand in hand

Månsson

2012

J Muscle Res Cell Motil

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This review describes the development towards actomyosin based nanodevices taking a starting point in pioneering studies in the 1990s based on conventional in vitro motility assays. References are given to parallel developments using the kinesin–microtubule motor system. The early developments focused on achieving cargo-transportation using actin filaments as cargo-loaded shuttles propelled by surface-adsorbed heavy meromyosin along micro- and nanofabricated channels. These efforts prompted extensive studies of surface–motor interactions contributing with new insights of general relevance in surface and colloid chemistry. As a result of these early efforts, a range of complex devices have now emerged, spanning applications in medical diagnostics, biocomputation and formation of complex nanostructures by self-organization. In addition to giving a comprehensive account of the developments towards real-world applications an important goal of the present review is to demonstrate important connections between the applied studies and fundamental biophysical studies of actomyosin and muscle function. Thus the manipulation of the motor proteins towards applications has resulted in new insights into methodological aspects of the in vitro motiliy assay. Other developments have advanced the understanding of the dynamic materials properties of actin filaments.

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Targeted Optimization of a Protein Nanomachine for Operation in Biohybrid Devices

Cited by 19 publications

References 37 publications

Small molecule-mediated refolding and activation of myosin motor function

Small molecule-mediated refolding and activation of myosin motor function

Diatrack particle tracking software: Review of applications and performance evaluation

Translational actomyosin research: fundamental insights and applications hand in hand

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