| 1. |
- Small, Lara S.R., et al.
(författare)
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Construction of a Chassis for a Tripartite Protein-Based Molecular Motor
- 2017
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Ingår i: ACS Synthetic Biology. - : American Chemical Society (ACS). - 2161-5063. ; 6:6, s. 1096-1102
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Tidskriftsartikel (refereegranskat)abstract
- Improving our understanding of biological motors, both to fully comprehend their activities in vital processes, and to exploit their impressive abilities for use in bionanotechnology, is highly desirable. One means of understanding these systems is through the production of synthetic molecular motors. We demonstrate the use of orthogonal coiled-coil dimers (including both parallel and antiparallel coiled coils) as a hub for linking other components of a previously described synthetic molecular motor, the Tumbleweed. We use circular dichroism, analytical ultracentrifugation, dynamic light scattering, and disulfide rearrangement studies to demonstrate the ability of this six-peptide set to form the structure designed for the Tumbleweed motor. The successful formation of a suitable hub structure is both a test of the transferability of design rules for protein folding as well as an important step in the production of a synthetic protein-based molecular motor.
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| 2. |
- Small, Lara S.R., et al.
(författare)
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The bar-hinge motor : A synthetic protein design exploiting conformational switching to achieve directional motility
- 2019
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Ingår i: New Journal of Physics. - : IOP Publishing. - 1367-2630. ; 21:1
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Tidskriftsartikel (refereegranskat)abstract
- One challenge to synthetic biology is to design functional machines from natural building blocks, from individual amino acids up to larger motifs such as the coiled coil. Here we investigate a novel bipedal motor concept, the Bar-Hinge Motor (BHM), a peptide-based motor capable of executing directed motion via externally controlled conformational switching between a straight bar and a V-shaped hinged form. Incorporating ligand-regulated binding to a DNA track and periodic control of ligand supply makes the BHM an example of a 'clocked walker'. Here, we employ a coarse-grained computational model for the BHM to assess the feasibility of a proposed experimental realization, with conformational switching regulated through the photoisomerization of peptide-bound azobenzene molecules. The results of numerical simulations using the model show that the incorporation of this conformational switch is necessary for the BHM to execute directional, rather than random, motion on a one-dimensional track. The power-stroke-driven directed motion is seen in the model even under conditions that underestimate the level of control we expect to be able to produce in the experimental realisation, demonstrating that this type of design should be an excellent vehicle for exploring the physics behind protein motion. By investigating its force-dependent dynamics, we show that the BHM is capable of directional motion against an applied load, even in the more relaxed conformational switching regimes. Thus, BHM appears to be an excellent candidate for a motor design incorporating a power stroke, enabling us to explore the ability of switchable coiled-coil designs to deliver power strokes within synthetic biology.
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| 3. |
- Boyle, Aimee L., et al.
(författare)
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Squaring the Circle in Peptide Assembly: From Fibers to Discrete Nanostructures by de Novo Design
- 2012
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Ingår i: Journal of the American Chemical Society. - : American Chemical Society (ACS). - 1520-5126 .- 0002-7863. ; 134:37, s. 15457-15467
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Tidskriftsartikel (refereegranskat)abstract
- The design of bioinspired nanostructures and materials of defined size and shape is challenging as it pushes our understanding of biomolecular assembly to its limits. In such endeavors, DNA is the current building block of choice because of its predictable and programmable self-assembly. The use of peptide- and protein-based systems, however, has potential advantages due to their more-varied chemistries, structures and functions, and the prospects for recombinant production through gene synthesis and expression. Here, we present the design and characterization of two complementary peptides programmed to form a parallel heterodimeric coiled coil, which we use as the building blocks for larger, supramolecular assemblies. To achieve the latter, the two peptides are joined via peptidic linkers of variable lengths to produce a range of assemblies, from flexible fibers of indefinite length, through large colloidal-scale assemblies, down to closed and discrete nanoscale objects of defined stoichiometry. We posit that the different modes of assembly reflect the interplay between steric constraints imposed by short linkers and the bulk of the helices, and entropic factors that favor the formation of many smaller objects as the linker length is increased. This approach, and the resulting linear and proteinogenic polypeptides, represents a new route for constructing complex peptide-based assemblies and biomaterials.
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| 4. |
- Bromley, Elizabeth H. C., et al.
(författare)
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The Tumbleweed: towards a synthetic protein motor
- 2009
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Ingår i: HFSP Journal. - : Informa UK Limited. - 1955-2068. ; 3:3, s. 204-212
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Tidskriftsartikel (refereegranskat)abstract
- Biomolecular motors have inspired the design and construction of artificial nanoscale motors and machines based on nucleic acids, small molecules, and inorganic nanostructures. However, the high degree of sophistication and efficiency of biomolecular motors, as well as their specific biological function, derives from the complexity afforded by protein building blocks. Here, we discuss a novel bottom-up approach to understanding biological motors by considering the construction of synthetic protein motors. Specifically, we present a design for a synthetic protein motor that moves along a linear track, dubbed the "Tumbleweed." This concept uses three discrete ligand-dependent DNA-binding domains to perform cyclically ligand-gated, rectified diffusion along a synthesized DNA molecule. Here we describe how de novo peptide design and molecular biology could be used to produce the Tumbleweed, and we explore the fundamental motor operation of such a design using numerical simulations. The construction of this and more sophisticated protein motors is an exciting challenge that is likely to enhance our understanding of the structure-function relationship in biological motors. [DOI: 10.2976/1.3111282]
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| 5. |
- Korosec, Chapin S., et al.
(författare)
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Motility of an autonomous protein-based artificial motor that operates via a burnt-bridge principle
- 2024
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Ingår i: Nature Communications. - : Springer Nature. - 2041-1723. ; 15:1
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Tidskriftsartikel (refereegranskat)abstract
- Inspired by biology, great progress has been made in creating artificial molecular motors. However, the dream of harnessing proteins – the building blocks selected by nature – to design autonomous motors has so far remained elusive. Here we report the synthesis and characterization of the Lawnmower, an autonomous, protein-based artificial molecular motor comprised of a spherical hub decorated with proteases. Its “burnt-bridge” motion is directed by cleavage of a peptide lawn, promoting motion towards unvisited substrate. We find that Lawnmowers exhibit directional motion with average speeds of up to 80 nm/s, comparable to biological motors. By selectively patterning the peptide lawn on microfabricated tracks, we furthermore show that the Lawnmower is capable of track-guided motion. Our work opens an avenue towards nanotechnology applications of artificial protein motors.
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| 6. |
- Kovacic, Suzana, et al.
(författare)
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Construction and Characterization of Kilobasepair Densely Labeled Peptide-DNA
- 2014
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Ingår i: Biomacromolecules. - : American Chemical Society (ACS). - 1526-4602 .- 1525-7797. ; 15:11, s. 4065-4072
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Tidskriftsartikel (refereegranskat)abstract
- Directed assembly of biocompatible materials benefits from modular building blocks in which structural organization is independent of introduced functional modifications. For soft materials, such modifications have been limited. Here, long DNA is successfully functionalized with dense decoration by peptides. Following introduction of alkyne-modified nucleotides into kilobasepair DNA, measurements of persistence length show that DNA mechanics are unaltered by the dense incorporation of alkynes (similar to 1 alkyne/2 bp) and after click-chemistry attachment of a tunable density of peptides. Proteolytic cleavage of densely tethered peptides (similar to 1 peptide/3 bp) demonstrates addressability of the functional groups, showing that this accessible approach to creating hybrid structures can maintain orthogonality between backbone mechanics and overlaid function. The synthesis and characterization of these hybrid constructs establishes the groundwork for their implementation in future applications, such as building blocks in modular approaches to a range of problems in synthetic biology.
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| 7. |
- Kovacic, Suzana, et al.
(författare)
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Design and Construction of a One- Dimensional DNA Track for an Artificial Molecular Motor
- 2012
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Ingår i: Journal of Nanomaterials. - : Hindawi Limited. - 1687-4129 .- 1687-4110.
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Tidskriftsartikel (refereegranskat)abstract
- DNA is a versatile heteropolymer that shows great potential as a building block for a diverse array of nanostructures. We present here a solution to the problem of designing and synthesizing a DNA-based nanostructure that will serve as the track along which an artificial molecular motor processes. This one-dimensional DNA track exhibits periodically repeating elements that provide specific binding sites for the molecular motor. Besides these binding elements, additional sequences are necessary to label specific regions within the DNA track and to facilitate track construction. Designing an ideal DNA track sequence presents a particular challenge because of the many variable elements that greatly expand the number of potential sequences from which the ideal sequence must be chosen. In order to find a suitable DNA sequence, we have adapted a genetic algorithm which is well suited for a large but sparse search space. This algorithm readily identifies long DNA sequences that include all the necessary elements to both facilitate DNA track construction and to present appropriate binding sites for the molecular motor. We have successfully experimentally incorporated the sequence identified by the algorithm into a long DNA track meeting the criteria for observation of the molecular motor's activity.
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| 8. |
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| 9. |
- Kovacic, Suzana, et al.
(författare)
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Design and Construction of the Lawnmower, An Artificial Burnt-Bridges Motor
- 2015
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Ingår i: IEEE Transactions on Nanobioscience. - 1558-2639. ; 14:3, s. 305-312
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Tidskriftsartikel (refereegranskat)abstract
- Molecular motors of the cell are protein-based, nanoscale machines, which use a variety of strategies to transduce chemical energy into mechanical work in the presence of a large thermal background. The design and construction of artificial molecular motors is one approach to better understand their basic physical principles. Here, we propose the concept of a protein-based, burnt-bridges ratchet, inspired by biological examples. Our concept, the lawnmower, utilizes protease blades to cleave peptide substrates, and uses the asymmetric substrate-product interface arising from productive cleavage to bias subsequent diffusion on the track (lawn). Following experimental screening to select a protease to act as the motor's blades, we chemically couple trypsin to quantum dots and demonstrate activity of the resulting lawnmower construct in solution. Accompanying Brownian dynamics simulations illustrate the importance for processivity of correct protease density on the quantum dot and spacing of substrates on the track. These results lay the groundwork for future tests of the protein-based lawnmower's motor performance characteristics.
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| 10. |
- Kuwada, Nathan J., et al.
(författare)
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Tuning the performance of an artificial protein motor
- 2011
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Ingår i: Physical Review E (Statistical, Nonlinear, and Soft Matter Physics). - 1539-3755. ; 84:3
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Tidskriftsartikel (refereegranskat)abstract
- The Tumbleweed (TW) is a concept for an artificial, tri-pedal, protein-based motor designed to move unidirectionally along a linear track by a diffusive tumbling motion. Artificial motors offer the unique opportunity to explore how motor performance depends on design details in a way that is open to experimental investigation. Prior studies have shown that TW's ability to complete many successive steps can be critically dependent on the motor's diffusional step time. Here, we present a simulation study targeted at determining how to minimize the diffusional step time of the TW motor as a function of two particular design choices: nonspecific motor-track interactions and molecular flexibility. We determine an optimal nonspecific interaction strength and establish a set of criteria for optimal molecular flexibility as a function of the nonspecific interaction. We discuss our results in the context of similarities to biological, linear stepping diffusive molecular motors with the aim of identifying general engineering principles for protein motors.
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