Actin-filament motion in the in vitro motility assay has a periodic component

deBeer, Evert L.; Sontrop, Annemiek M.A.T.A.; Kellermayer, Miklós; Galambos, Csaba; Pollack, Gerald H.

doi:10.1002/(sici)1097-0169(1997)38:4<341::aid-cm4>3.0.co;2-6

Cited by 15 publications

(1 citation statement)

References 30 publications

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“…The filament motion displayed acceleration/deceleration phases throughout the entire time course of the movement. This periodicity, which has been reported earlier [49,50], is probably due to the heterogeneity of the HMM molecules coated on the glass surface, although other explanations such as intra-actin cooperativity have also been proposed.…”

Section: T800 Accelerates In Vitro Motility Of the Reconstituted Thinsupporting

confidence: 77%

Regulation of the actin–myosin interaction by titin

Niederländer

Raynaud

Astier

et al. 2004

European Journal of Biochemistry

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Titin is known to interact with actin thin filaments within the I-band region of striated muscle sarcomeres. In this study, we have used a titin fragment of 800 kDa (T800) purified from striated skeletal muscle to measure the effect of this interaction on the functional properties of the actinmyosin complex. MALDI-TOF MS revealed that T800 contains the entire titin PEVK (Pro, Glu, Val, Lys-rich) 1 domain. In the presence of tropomyosin-troponin, T800 increased the sliding velocity (both average and maximum values) of actin filaments on heavy-meromyosin (HMM)-coated surfaces and dramatically decreased the number of stationary filaments. These results were correlated with a 30% reduction in actin-activated HMM ATPase activity and with an inhibition of HMM binding to actin N-terminal residues as shown by chemical cross-linking. At the same time, T800 did not affect the efficiency of the Ca 2+ -controlled on/off switch, nor did it alter the overall binding energetics of HMM to actin, as revealed by cosedimentation experiments. These data are consistent with a competitive effect of PEVK domain-containing T800 on the electrostatic contacts at the actin-HMM interface. They also suggest that titin may participate in the regulation of the active tension generated by the actin-myosin complex.Keywords: ATPase; chemical cross-linking; mass spectrometry; motility assay; muscle contraction.Titin is the largest known protein, containing more than 38 000 residues in its longest human striated muscle isoform. It represents the third most abundant component of vertebrate striated muscle, after myosin and actin, and is also present in smooth muscle and nonmuscle cells (recently reviewed in [1,2]). The importance of intact titin for normal muscle function has been demonstrated in vitro [3][4][5], as well as in vivo through its implication in muscular dystrophies such as dilated cardiomyopathies and Udd's tibial muscular dystrophy (reviewed in [6]).In striated muscle, titin is involved in several fundamental processes, including sarcomere assembly, possibly in thick filament length control [4,[7][8][9], maintenance of the sarcomeric structure, muscle elasticity and passive tension development [10][11][12]. These functions are related to three main structural properties of the protein: titin spans half a sarcomere, from the Z disks to the M line (connecting the Z disks to myosin thick filaments), it contains subdomains that confer unusual elastic properties, and it interacts with several protein partners such as myosin, actin, M protein, C protein, MURF-1, calpain 3, myomesin, a-actinin, nebulin, telethonin and obscurin.The elastic domains are made of tandemly arranged immunoglobulin (Ig)-like domains and a unique PEVK domain (Pro, Glu, Val, Lys-rich) whose size depends on the muscle fibre isotype. Specific structural properties and mechanical force/extension measurements made on muscle fibres or at the single molecule level suggest that the tandem Ig-and PEVK-domains are two elements of differential stiffness that function as a two-...

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Section: T800 Accelerates In Vitro Motility Of the Reconstituted Thinsupporting

confidence: 77%

Regulation of the actin–myosin interaction by titin

Niederländer

Raynaud

Astier

et al. 2004

European Journal of Biochemistry

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The Relation of Different-Scale Membrane Processes Under Nitric Oxide Influence

et al. 2005

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Rhythmic activity, synaptic transmission and propagation of excitation depend on the processes that occur at different cellular levels. Modulation of neuron activity results from the changes of plasma membrane and organelles properties. We aimed to study the relationship between changes of the ion channels activity, membrane microviscosity and amount of bound Ca(2+) under the influence of nitric oxide (NO). We also investigated the effect of NO on the refractive index of neurons. We have shown that NO activates voltage-dependent K(K(V))-channels and leads to the decrease in the amount of membrane-bound and stored Ca(2+). NO causes a decrease in the microviscosity of the membrane of cytosomes and altered refractive index of neurones. The latter can result from the modification of plasma membrane physico-chemical properties and structural changes in the cytoplasm (vesicles movement, reorganisation of cytoskeleton, etc.). It is suggested that long-term modifications of the physico-chemical properties of membranes and reorganisation in the cytoplasm affect neuron activity and signal transmission.

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Up-and-down movement of a sliding actin filament in the in vitro motility assay

2011

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a b s t r a c tWe observed a three-dimensional up-and-down movement of an actin filament sliding on heavy meromyosin (HMM) molecules in an in vitro motility assay. The up-and-down movement occurred along the direction perpendicular to the planar glass plane on which the filament demonstrated a sliding movement. The height length of the up-and-down movement was measured by monitoring the extent of diminishing fluorescent emission from the marker attached to the filament in the evanescent field of attenuation. The height lengths whose distribution exhibits a local maximum were found around the two values, 150 nm and 90 nm, separately. This undulating three-dimensional movement of an actin filament suggests that the interactions between myosin (HMM) molecules and the actin filament may temporally be modulated during its sliding movement.

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Actin-filament motion in the in vitro motility assay has a periodic component

Cited by 15 publications

References 30 publications

Regulation of the actin–myosin interaction by titin

Regulation of the actin–myosin interaction by titin

The Relation of Different-Scale Membrane Processes Under Nitric Oxide Influence

Up-and-down movement of a sliding actin filament in the in vitro motility assay

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