scholarly journals The molecular basis of frictional loads in the in vitro motility assay with applications to the study of the loaded mechanochemistry of molecular motors

Cytoskeleton ◽  
2010 ◽  
Vol 67 (5) ◽  
pp. 273-285 ◽  
Author(s):  
Michael J. Greenberg ◽  
Jeffrey R. Moore
Keyword(s):  
Molecular Motors ◽  
Molecular Basis ◽  
Motility Assay ◽  
In Vitro Motility Assay ◽  
In Vitro Motility

scholarly journals An automated in vitro motility assay for high-throughput studies of molecular motors

Lab on a Chip ◽  
2018 ◽  
Vol 18 (20) ◽  
pp. 3196-3206 ◽  
Author(s):  
Till Korten ◽  
Elena Tavkin ◽  
Lara Scharrel ◽  
Vandana Singh Kushwaha ◽  
Stefan Diez
Keyword(s):  
High Throughput ◽  
Molecular Motors ◽  
Lab On A Chip ◽  
Force Generation ◽  
Motility Assay ◽  
In Vitro Motility Assay ◽  
In Vitro Motility ◽  
Cargo Transport

Molecular motors, essential to force-generation and cargo transport within cells, are invaluable tools for powering nanobiotechnological lab-on-a-chip devices.

The effects of phosphate and acidosis on regulated thin-filament velocity in an in vitro motility assay

2012 ◽  
Vol 113 (9) ◽  
pp. 1413-1422 ◽  
Author(s):  
Edward P. Debold ◽  
Thomas J. Longyear ◽  
Matthew A. Turner
Keyword(s):  
Molecular Basis ◽  
Bound States ◽  
Contractile Activity ◽  
Motility Assay ◽  
Shortening Velocity ◽  
In Vitro Motility Assay ◽  
In Vitro Motility ◽  
Specific Step ◽  
Regulated Thin Filament

Muscle fatigue from intense contractile activity is thought to result, in large part, from the accumulation of inorganic phosphate (Pi) and hydrogen ions (H+) acting to directly inhibit the function of the contractile proteins; however, the molecular basis of this process remain unclear. We used an in vitro motility assay and determined the effects of elevated H+ and Pi on the ability of myosin to bind to and translocate regulated actin filaments (RTF) to gain novel insights into the molecular basis of fatigue. At saturating Ca++, acidosis depressed regulated filament velocity ( VRTF) by ∼90% (6.2 ± 0.3 vs. 0.5 ± 0.2 μm/s at pH 7.4 and 6.5, respectively). However, the addition of 30 mM Pi caused VRTF to increase fivefold, from 0.5 ± 0.2 to 2.6 ± 0.3 μm/s at pH 6.5. Similarly, at all subsaturating Ca++ levels, acidosis slowed VRTF, but the addition of Pi significantly attenuated this effect. We also manipulated the [ADP] in addition to the [Pi] to probe which specific step(s) of cross-bridge cycle of myosin is affected by elevated H+. The findings are consistent with acidosis slowing the isomerization step between two actomyosin ADP-bound states. Because the state before this isomerization is most vulnerable to Pi rebinding, and the associated detachment from actin, this finding may also explain the Pi-induced enhancement of VRTF at low pH. These results therefore may provide a molecular basis for a significant portion of the loss of shortening velocity and possibly muscular power during fatigue.

Caged FEDA-ATP: a new tool in the measurement of ATP turnover during the in vitro motility assay

1995 ◽  
Vol 23 (3) ◽  
pp. 401S-401S ◽  
Author(s):  
Daren S. Jeffreys ◽  
Robert J. Eaton ◽  
Clive R. Bagshaw
Keyword(s):  
Motility Assay ◽  
In Vitro Motility Assay ◽  
In Vitro Motility ◽  
Atp Turnover

scholarly journals Determination of the Maximum Velocity of Filaments in the in vitro Motility Assay

2019 ◽  
Vol 10 ◽  
Author(s):  
Nasrin Bopp ◽  
Lisa-Mareike Scheid ◽  
Rainer H. A. Fink ◽  
Karl Rohr
Keyword(s):  
Maximum Velocity ◽  
Motility Assay ◽  
In Vitro Motility Assay ◽  
In Vitro Motility

Sliding velocity of isolated rabbit cardiac myosin correlates with isozyme distribution

1992 ◽  
Vol 263 (2) ◽  
pp. H464-H472 ◽  
Author(s):  
H. Yamashita ◽  
S. Sugiura ◽  
T. Serizawa ◽  
T. Sugimoto ◽  
M. Iizuka ◽  
...  
Keyword(s):  
Atpase Activity ◽  
Myosin Heavy Chain ◽  
Heavy Chain ◽  
Motility Assay ◽  
Cardiac Myosin ◽  
Sliding Velocity ◽  
In Vitro Motility Assay ◽  
In Vitro Motility ◽  
Actin Cables

To investigate the relationship between the mechanical and biochemical properties of cardiac myosin, the sliding velocity of isolated cardiac myosin obtained from both euthyroid and hyperthyroid rabbits on actin cables was measured with an in vitro motility assay system. Ten rabbits (T) were treated with L-thyroxine to induce hyperthyroidism, and eight nontreated animals (N) were used as controls. Myosin was purified from the left ventricles of anesthetized animals. Myosin isozyme content was analyzed by the pyrophosphate gel electrophoresis method, and myosin adenosinetriphosphatase (ATPase) activity was determined on the same sample. Long well-organized actin cables of green algae, Nitellopsis, were used in the in vitro motility assay. Small latex beads were coated with purified cardiac myosin and introduced onto the Nitellopsis actin cables. Active unidirectional movement of the beads on the actin cables was observed under a photomicroscope, and the velocity was measured. The velocity was dependent on ATP concentrations, and the optimal pH for bead movement was approximately 7.0-7.5. The mean velocity was higher in T than in N (0.66 +/- 0.12 vs. 0.32 +/- 0.09 micron/s, P less than 0.01). Both Ca(2+)-activated ATPase activity and the percentage of alpha-myosin heavy chain were also higher in T than in N (0.691 +/- 0.072 vs. 0.335 +/- 0.072 microM Pi.mg-1.min-1, P less than 0.01, and 79 +/- 12 vs. 26 +/- 7%, P less than 0.01, respectively). The velocity of myosin closely correlated with both Ca(+2)-activated myosin ATPase activity (r = 0.87, P less than 0.01) and the percentage of alpha-myosin heavy chain (r = 0.87, P less than 0.01).

Path Reconstruction as a Tool for Actin Filament Speed Determination in the in Vitro Motility Assay

1999 ◽  
Vol 273 (1) ◽  
pp. 12-19 ◽  
Author(s):  
W. Hamelink ◽  
J.G. Zegers ◽  
B.W. Treijtel ◽  
T. Blangé
Keyword(s):  
Actin Filament ◽  
Motility Assay ◽  
In Vitro Motility Assay ◽  
In Vitro Motility
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