Velocity-dependent Actomyosin ATPase Cycle Revealed by in Vitro Motility Assay with Kinetic Analysis

Overview

Journal Biophys J

Publisher Cell Press

Specialty Biophysics

Date 2012 Sep 6

PMID 22947932

Citations 5

Authors

Masaaki K Sato

Takashi Ishihara

Hiroto Tanaka

Akihiko Ishijima

Yuichi Inoue

Affiliations

Soon will be listed here.

Abstract

The actomyosin interaction plays a key role in a number of cellular functions. Single-molecule measurement techniques have been developed to study the mechanism of the actomyosin contractile system. However, the behavior of isolated single molecules does not always reflect that of molecules in a complex system such as a muscle fiber. Here, we developed a simple method for studying the kinetic parameters of the actomyosin interaction using small numbers of molecules. This approach does not require the specialized equipment needed for single-molecule measurements, and permits us to observe behavior that is more similar to that of a complex system. Using an in vitro motility assay, we examined the duration of continuous sliding of actin filaments on a sparsely distributed heavy meromyosin-coated surface. To estimate the association rate constant of the actomyosin motile system, we compared the distribution of experimentally obtained duration times with a computationally simulated distribution. We found that the association rate constant depends on the sliding velocity of the actin filaments. This technique may be used to reveal new aspects of the kinetics of various motor proteins in complex systems.

Citing Articles

Cell Behavioral Dynamics as a Cue in Optimizing Culture Stabilization in the Bioprocessing of Pluripotent Stem Cells.

Thanuthanakhun N, Kim M, Kino-Oka M Bioengineering (Basel). 2022; 9(11).

PMID: 36354580 PMC: 9687444. DOI: 10.3390/bioengineering9110669.

Kallenberger S, Unger A, Legewie S, Lymperopoulos K, Klingmuller U, Eils R PLoS Comput Biol. 2017; 13(9):e1005779.

PMID: 28945754 PMC: 5659801. DOI: 10.1371/journal.pcbi.1005779.

A small-molecule modulator of cardiac myosin acts on multiple stages of the myosin chemomechanical cycle.

Kawas R, Anderson R, Ingle S, Song Y, Sran A, Rodriguez H J Biol Chem. 2017; 292(40):16571-16577.

PMID: 28808052 PMC: 5633120. DOI: 10.1074/jbc.M117.776815.

Maximum limit to the number of myosin II motors participating in processive sliding of actin.

Rastogi K, Puliyakodan M, Pandey V, Nath S, Elangovan R Sci Rep. 2016; 6:32043.

PMID: 27554800 PMC: 4995457. DOI: 10.1038/srep32043.

Single turnovers of fluorescent ATP bound to bipolar myosin filament during actin filaments sliding.

Maruta T, Kobatake T, Okubo H, Chaen S Biophysics (Nagoya-shi). 2016; 9:13-20.

PMID: 27493536 PMC: 4629674. DOI: 10.2142/biophysics.9.13.

References

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Yanagida T, Nakase M, Nishiyama K, Oosawa F . Direct observation of motion of single F-actin filaments in the presence of myosin. Nature. 1984; 307(5946):58-60. DOI: 10.1038/307058a0. View

Molloy J, Burns J, Sparrow J, Tregear R, Kendrick-Jones J, White D . Single-molecule mechanics of heavy meromyosin and S1 interacting with rabbit or Drosophila actins using optical tweezers. Biophys J. 1995; 68(4 Suppl):298S-303S; 303S-305S. PMC: 1281954. View

Ishijima A, Doi T, Sakurada K, Yanagida T . Sub-piconewton force fluctuations of actomyosin in vitro. Nature. 1991; 352(6333):301-6. DOI: 10.1038/352301a0. View

Siddique M, Mogami G, Miyazaki T, Katayama E, Uyeda T, Suzuki M . Cooperative structural change of actin filaments interacting with activated myosin motor domain, detected with copolymers of pyrene-labeled actin and acto-S1 chimera protein. Biochem Biophys Res Commun. 2005; 337(4):1185-91. DOI: 10.1016/j.bbrc.2005.09.159. View

Nyitrai M, Geeves M . Adenosine diphosphate and strain sensitivity in myosin motors. Philos Trans R Soc Lond B Biol Sci. 2005; 359(1452):1867-77. PMC: 1693474. DOI: 10.1098/rstb.2004.1560. View

White H, Taylor E . Energetics and mechanism of actomyosin adenosine triphosphatase. Biochemistry. 1976; 15(26):5818-26. DOI: 10.1021/bi00671a020. View

Steffen W, Smith D, Simmons R, Sleep J . Mapping the actin filament with myosin. Proc Natl Acad Sci U S A. 2001; 98(26):14949-54. PMC: 64964. DOI: 10.1073/pnas.261560698. View

Svoboda K, Schmidt C, Schnapp B, Block S . Direct observation of kinesin stepping by optical trapping interferometry. Nature. 1993; 365(6448):721-7. DOI: 10.1038/365721a0. View

10.

Kron S, Toyoshima Y, Uyeda T, Spudich J . Assays for actin sliding movement over myosin-coated surfaces. Methods Enzymol. 1991; 196:399-416. DOI: 10.1016/0076-6879(91)96035-p. View

11.

Lymn R, Taylor E . Mechanism of adenosine triphosphate hydrolysis by actomyosin. Biochemistry. 1971; 10(25):4617-24. DOI: 10.1021/bi00801a004. View

12.

Spudich J, Watt S . The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971; 246(15):4866-71. View

13.

Shingyoji C, Higuchi H, Yoshimura M, Katayama E, Yanagida T . Dynein arms are oscillating force generators. Nature. 1998; 393(6686):711-4. DOI: 10.1038/31520. View

14.

Tokuraku K, Kurogi R, Toya R, Uyeda T . Novel mode of cooperative binding between myosin and Mg2+ -actin filaments in the presence of low concentrations of ATP. J Mol Biol. 2008; 386(1):149-62. DOI: 10.1016/j.jmb.2008.12.008. View

15.

Huxley H, Brown W . The low-angle x-ray diagram of vertebrate striated muscle and its behaviour during contraction and rigor. J Mol Biol. 1967; 30(2):383-434. DOI: 10.1016/s0022-2836(67)80046-9. View

16.

Sleep J, Hutton R . Actin mediated release of ATP from a myosin-ATP complex. Biochemistry. 1978; 17(25):5423-30. DOI: 10.1021/bi00618a016. View

17.

Stein L, Schwarz Jr R, Chock P, Eisenberg E . Mechanism of actomyosin adenosine triphosphatase. Evidence that adenosine 5'-triphosphate hydrolysis can occur without dissociation of the actomyosin complex. Biochemistry. 1979; 18(18):3895-909. DOI: 10.1021/bi00585a009. View

18.

Schnitzer M, Block S . Kinesin hydrolyses one ATP per 8-nm step. Nature. 1997; 388(6640):386-90. DOI: 10.1038/41111. View

19.

Toyoshima Y, Toyoshima C, Spudich J . Bidirectional movement of actin filaments along tracks of myosin heads. Nature. 1989; 341(6238):154-6. DOI: 10.1038/341154a0. View

20.

Bowater R, Webb M, Ferenczi M . Measurement of the reversibility of ATP binding to myosin in calcium-activated skinned fibers from rabbit skeletal muscle. Oxygen exchange between water and ATP released to the solution. J Biol Chem. 1989; 264(13):7193-201. View