|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 276, Issue 43, 39812-39818, October 26, 2001
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the Brain myosin-Va consists of two heavy chains,
each containing a neck domain with six tandem IQ motifs that
bind four to five calmodulins and one to two essential light chains.
Previous studies demonstrated that myosin-Va exhibits an unusually high
affinity for F-actin in the presence of ATP and that its MgATPase
activity is stimulated by micromolar Ca2+ in a highly
cooperative manner. We demonstrate here that Ca2+ also
induces myosin-Va binding to and cosedimentation with F-actin in the
presence of ATP in a similar cooperative manner and calcium concentration range as that observed for the ATPase activity. Neither
hydrolysis of ATP nor buildup of ADP was required for Ca2+-induced cosedimentation. The Ca2+-induced
binding was inhibited by low temperature or by 0.6 M NaCl,
but not by 1% Triton X-100. Tight binding between myosin-Va and
pyrene-labeled F-actin in the presence of ATP and Ca2+ was
also detected by quenching of the pyrene fluorescence. Negatively stained preparations of actomyosin-Va under Ca2+-induced
binding conditions showed tightly packed F-actin bundles cross-linked
by myosin-Va. Our data demonstrate that high affinity binding of
myosin-Va and F-actin in the presence of ATP or
5'-O-(thiotriphosphate) is induced by micromolar
concentrations of Ca2+. Since Ca2+ regulates
both the actin binding properties and actin-activated ATPase of
myosin-Va over the same concentration range, we suggest that the
calcium signal may regulate the mechanism of processivity of myosin Va.
High Affinity Binding of Brain Myosin-Va to F-actin Induced by
Calcium in the Presence of ATP*
§¶,
,, and§§
Department of Cellular and Molecular
Biology, Faculdade de Medicina de Ribeirão Preto, Universidade de
São Paulo, Ribeirão Preto, SP, Brazil, 14049-900, the
§ Marine Biological Laboratory, Woods Hole, Massachusetts
02543, the ** Department of Cell & Molecular Biology,
Northwestern University Medical School, Chicago, Illinois 60611, and
the Departments of MCD-Biology, Cell
Biology, and Pathology, Yale University, New
Haven, Connecticut 06511
*
This work was supported by grants (to R. E. L.)
from the Fundação de Amparo à Pesquisa do Estado de
São Paulo (FAPESP), Programa de Apoio ao Desenvolvimento
Científico e Tecnológico (PADCT), Conselho Nacional de
Desenvolvimento Científico e Tecnológico (CNPq), and
Fundação Apoio ao Ensino, Pesquisa e Assistência do
Hospital das Clínicas da Faculdade de Medicina de
Ribeirão Preto (FAEPA) and by National Institutes of Health
Grants DK 25387 and DK 55389 (to M. S. M.). A portion of
these studies was conducted in the Physiology Course (1998), Marine
Biological Laboratory, Woods Hole, MA.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Held an undergraduate stipend from FAPESP.
§§
To whom correspondence should be addressed. Tel.: 55-16-602-3319;
Fax: 55-16-633-1786; E-mail: relarson@fmrp.usp.br.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  A. O. Olivares, W. Chang, M. S. Mooseker, D. D. Hackney, and E. M. De La Cruz The Tail Domain of Myosin Va Modulates Actin Binding to One Head J. Biol. Chem., October 20, 2006; 281(42): 31326 - 31336. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Yi, D. Weaver, and G. Hajnoczky Control of mitochondrial motility and distribution by the calcium signal: a homeostatic circuit J. Cell Biol., November 22, 2004; 167(4): 661 - 672. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. N. Krementsov, E. B. Krementsova, and K. M. Trybus Myosin V: regulation by calcium, calmodulin, and the tail domain J. Cell Biol., March 15, 2004; 164(6): 877 - 886. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Wang, K. Thirumurugan, W. F. Stafford, J. A. Hammer III, P. J. Knight, and J. R. Sellers Regulated Conformation of Myosin V J. Biol. Chem., January 23, 2004; 279(4): 2333 - 2336. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. T. Cao, W. Chang, S. E. Masters, and M. S. Mooseker Myosin-Va Binds to and Mechanochemically Couples Microtubules to Actin Filaments Mol. Biol. Cell, January 1, 2004; 15(1): 151 - 161. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.-C. Chen-Hwang, H.-R. Chen, M. Elzinga, and Y.-W. Hwang Dynamin Is a Minibrain Kinase/Dual Specificity Yak1-related Kinase 1A Substrate J. Biol. Chem., May 10, 2002; 277(20): 17597 - 17604. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. L. Post, M. J. Tyska, C. B. O'Connell, K. Johung, A. Hayward, and M. S. Mooseker Myosin-IXb Is a Single-headed and Processive Motor J. Biol. Chem., March 29, 2002; 277(14): 11679 - 11683. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |