Skip Navigation



Plant and Cell Physiology Advance Access published online on May 15, 2007
Plant and Cell Physiology, doi:10.1093/pcp/pcm054
This Article
Right arrow Advance Access manuscript (PDF)
Right arrow All Versions of this Article:
48/6/886    most recent
pcm054v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Hachikubo, Y.
Right arrow Articles by Yamamoto, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hachikubo, Y.
Right arrow Articles by Yamamoto, K.
Agricola
Right arrow Articles by Hachikubo, Y.
Right arrow Articles by Yamamoto, K.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Enzymatic activity and motility of recombinant Arabidopsis myosin XI, MYA1

You Hachikubo1, Kohji Ito1, John Schiefelbein2, Dietmar J. Manstein3 and Keiichi Yamamoto1,*

1 Department of Biology, Chiba University, Inageku, Chiba 263-8522, Japan
2 Department of Molecular, Cellular, and Developmental Biology, University of Michigan, MI, USA
3 Institute for Biophysical Chemistry, Hannover Medical School, Hannover, Germany

*To whom all correspondence should be addressed: Keiichi Yamamoto. Department of Biology, Chiba University, Inageku, Chiba 263-8522, Japan. Phone: +81 43 290 2809, FAX: +81 43 290 2809, E-mail: yamamoto{at}bio.s.chiba-u.ac.jp


  Abstract

We expressed recombinant Arabidopsis myosin XI (MYA1) in which the motor domain of MYA1 was connected to an artificial lever arm composed of triple helical repeats of Dictyostelium {alpha}-actinin to understand its motor activity and intracellular function. The Vmax and Kactin of the actin-activated Mg2+ ATPase activity of the recombinant MYA1 were 50.7 Pi head-1 s-1 and 30.2 µM, respectively at 25 °C. The recombinant MYA1 could translocate actin filament at the maximum velocity of 1.8 µm s-1 at 25 °C in the in vitro motility assay. The value corresponded to the motility of 3.2 µm s-1 for native MYA1 if we consider the difference in the lever arm length and this value was very close to the velocity of cytoplasmic streaming in Arabidopsis hypocotyl epidermal cells. Extent of inhibition by ADP on the motility of MYA1 was similar to that of the well-known processive motor, myosin V, suggesting that MYA1 is a processive motor. Dissociation rate of actin-MYA1-ADP complex induced by ATP (73.5 s-1) and the Vmax value of the actin-activated Mg2+ ATPase activity revealed that MYA1 stays in the actin bound states for about 70 % of its mechanochemical cycle time. This high ratio of actin bound states is also a characteristic of processive motor. Our results strongly suggest that MYA1 is a processive motor and involved in vesicle transport and/or in cytoplasmic streaming.

Keywords: Arabidopsis - myosin - ATPase - processivity - motility - MYA1


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 J Exp BotHome page
E. Yokota, S. Ueda, K. Tamura, H. Orii, S. Uchi, S. Sonobe, I. Hara-Nishimura, and T. Shimmen
An isoform of myosin XI is responsible for the translocation of endoplasmic reticulum in tobacco cultured BY-2 cells
J. Exp. Bot., January 1, 2009; 60(1): 197 - 212.
[Abstract] [Full Text] [PDF]

Home page
 J Exp BotHome page
I. A. Sparkes, N. A. Teanby, and C. Hawes
Truncated myosin XI tail fusions inhibit peroxisome, Golgi, and mitochondrial movement in tobacco leaf epidermal cells: a genetic tool for the next generation
J. Exp. Bot., June 1, 2008; 59(9): 2499 - 2512.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
G. Tsiavaliaris, S. Fujita-Becker, U. Durrwang, R. P. Diensthuber, M. A. Geeves, and D. J. Manstein
Mechanism, Regulation, and Functional Properties of Dictyostelium Myosin-1B
J. Biol. Chem., February 22, 2008; 283(8): 4520 - 4527.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.