Biased Brownian motion mechanism for processivity and directionality of single-headed myosin-VI
Soft Biosystem Group, Laboratories for Nanobiology, Graduate School of Frontier Biosciences, Osaka University, 1-3,Yamadaoka, Suita, Osaka 565-871, Japan.Biosystems (Impact Factor: 1.55). 07/2008; 93(1-2):39-47. DOI: 10.1016/j.biosystems.2008.03.013
Conventional form to function as a vesicle transporter is not a 'single molecule' but a coordinated 'two molecules'. The coordinated two molecules make it complicated to reveal its mechanism. To overcome the difficulty, we adopted a single-headed myosin-VI as a model protein. Myosin-VI is an intracellular vesicle and organelle transporter that moves along actin filaments in a direction opposite to most other known myosin classes. The myosin-VI was expected to form a dimer to move processively along actin filaments with a hand-over-hand mechanism like other myosin organelle transporters. However, wild-type myosin-VI was demonstrated to be monomer and single-headed, casting doubt on its processivity. Using single molecule techniques, we show that green fluorescent protein (GFP)-fused single-headed myosin-VI does not move processively. However, when coupled to a 200 nm polystyrene bead (comparable to an intracellular vesicle in size) at a ratio of one head per bead, single-headed myosin-VI moves processively with large (40 nm) steps. Furthermore, we found that a single-headed myosin-VI-bead complex moved more processively in a high-viscous solution (40-fold higher than water) similar to cellular environment. Because diffusion of the bead is 60-fold slower than myosin-VI heads alone in water, we propose a model in which the bead acts as a diffusional anchor for the myosin-VI, enhancing the head's rebinding following detachment and supporting processive movement of the bead-monomer complex. This investigation will help us understand how molecular motors utilize Brownian motion in cells.
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ABSTRACT: It is puzzling that in spite of its single-headed structure, myosin-IX can move processively along actin. Here, based on the experimental evidence that the strong binding of myosin to actin in rigor state induces structural changes to several local actin monomers, a Brownian ratchet model is proposed to describe this processive movement. In the model, the actin plays an active role in the motility of single-headed myosin, in contrast to the common belief that the actin acts only as a passive track for the motility of the myosin. The unidirectional movement is due to both the asymmetric periodic potential of the myosin interacting with actin and the forward Stokes force induced by the relative rotation of the neck domain to the motor domain, while the processivity is determined by the binding affinity of the myosin for actin in ATP state. This gives a good explanation to the high processivity of myosin-IX, which results from its high binding affinity for actin in ATP state due to the presence of unique loop 2 insertion or N-terminal extension. The experimental results on the motility of myosin-IX such as the step size, large forward/backward stepping ratio, run length, stall force, etc, are explained well.Biophysical chemistry 09/2010; 151(1-2):71-80. DOI:10.1016/j.bpc.2010.05.007 · 1.99 Impact Factor
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ABSTRACT: Kinesin is a family of molecular motors that move unidirectionally along microtubules (MT) using ATP hydrolysis free energy. In the family, the conventional two-headed kinesin was experimentally characterized to move unidirectionally through "walking" in a hand-over-hand fashion by coordinated motions of the two heads. Interestingly a single-headed kinesin, a truncated KIF1A, still can generate a biased Brownian movement along MT, as observed by in vitro single molecule experiments. Thus, KIF1A must use a different mechanism from the conventional kinesin to achieve the unidirectional motions. Based on the energy landscape view of proteins, for the first time, we conducted a set of molecular simulations of the truncated KIF1A movements over an ATP hydrolysis cycle and found a mechanism exhibiting and enhancing stochastic forward-biased movements in a similar way to those in experiments. First, simulating stand-alone KIF1A, we did not find any biased movements, while we found that KIF1A with a large friction cargo-analog attached to the C-terminus can generate clearly biased Brownian movements upon an ATP hydrolysis cycle. The linked cargo-analog enhanced the detachment of the KIF1A from MT. Once detached, diffusion of the KIF1A head was restricted around the large cargo which was located in front of the head at the time of detachment, thus generating a forward bias of the diffusion. The cargo plays the role of a diffusional anchor, or cane, in KIF1A "walking."PLoS Computational Biology 02/2013; 9(2):e1002907. DOI:10.1371/journal.pcbi.1002907 · 4.62 Impact Factor
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