| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Biophysical Journal 68: 2444-2453 (1995)
© 1995 the Biophysical Society
Department of Pathology, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
ABSTRACT
We have analyzed the dependence of actin filament sliding movement on the mode of myosin attachment to surfaces. Monoclonal antibodies (mAbs) that bind to three distinct sites were used to tether myosin to nitrocellulose-coated glass. One antibody reacts with an epitope on the regulatory light chain (LC2) located at the head-rod junction. The other two react with sites in the rod domain, one in the S2 region near the S2-LMM hinge, and the other at the C terminus of the myosin rod. This method of attachment provides a means of controlling the flexibility and density of myosin on the surface. Fast skeletal muscle myosin monomers were bound to the surfaces through the specific interaction with these mAbs, and the sliding movement of fluorescently labeled actin filaments was analyzed by video microscopy. Each of these antibodies produced stable myosin-coated surfaces that supported uniform motion of actin over the course of several hours. Attachment of myosin through the anti-S2 and anti-LMM mAbs yielded significantly higher velocities (10 microns/s at 30 degrees C) than attachment through anti-LC2 (4-5 microns/s at 30 degrees C). For each antibody, we observed a characteristic value of the myosin density for the onset of F-actin motion and a second critical density for velocity saturation. The specific mode of attachment influences the velocity of actin filaments and the characteristic surface density needed to support movement.
This article has been cited by other articles:
 |
|
 |
|
  Q. Wang, C. L. Moncman, and D. A. Winkelmann Mutations in the motor domain modulate myosin activity and myofibril organization J. Cell Sci., October 15, 2003; 116(20): 4227 - 4238. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Chow, R. Srikakulam, Y. Chen, and D. A. Winkelmann Folding of the Striated Muscle Myosin Motor Domain J. Biol. Chem., September 20, 2002; 277(39): 36799 - 36807. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Badoual, F. Julicher, and J. Prost Bidirectional cooperative motion of molecular motors PNAS, May 14, 2002; 99(10): 6696 - 6701. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. M. Gordon, E. Homsher, and M. Regnier Regulation of Contraction in Striated Muscle Physiol Rev, April 1, 2000; 80(2): 853 - 924. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. G. Gillespie, S. K. H. Gillespie, J. A. Mercer, K. Shah, and K. M. Shokat Engineering of the Myosin-Ibeta Nucleotide-binding Pocket to Create Selective Sensitivity to N6-modified ADP Analogs J. Biol. Chem., October 29, 1999; 274(44): 31373 - 31381. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Srikakulam and D. A. Winkelmann Myosin II Folding Is Mediated by a Molecular Chaperonin J. Biol. Chem., September 17, 1999; 274(38): 27265 - 27273. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Kato, T. Nishizaka, T. Iga, K. Kinosita Jr., and S.'i. Ishiwata Imaging of thermal activation of actomyosin motors PNAS, August 17, 1999; 96(17): 9602 - 9606. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Gutsche-Perelroizen, J. Lepault, A. Ott, and M.-F. Carlier Filament Assembly from Profilin-Actin J. Biol. Chem., March 5, 1999; 274(10): 6234 - 6243. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. L. Sweeney, S. S. Rosenfeld, F. Brown, L. Faust, J. Smith, J. Xing, L. A. Stein, and J. R. Sellers Kinetic Tuning of Myosin via a Flexible Loop Adjacent to the Nucleotide Binding Pocket J. Biol. Chem., March 13, 1998; 273(11): 6262 - 6270. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Post, G. Bokoch, and M. Mooseker Human myosin-IXb is a mechanochemically active motor and a GAP for rho J. Cell Sci., January 4, 1998; 111(7): 941 - 950. [Abstract] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
