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  Transport efficiency of membrane-anchored kinesin-1 motors depends on motor density and diffusivity

Grover, R., Fischer, J., Schwarz, F. W., Walter, W. J., Schwille, P., & Diez, S. (2016). Transport efficiency of membrane-anchored kinesin-1 motors depends on motor density and diffusivity. Proceedings of the National Academy of Sciences of the United States of America, 113(46), E7185-E7193. doi:10.1073/pnas.1611398113.

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http://www.pnas.org/content/113/46/E7185 (Publisher version)
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 Creators:
Grover, Rahul1, Author
Fischer, Janine1, Author
Schwarz, Friedrich W.1, Author
Walter, Wilhelm J.1, Author
Schwille, Petra2, Author           
Diez, Stefan1, Author
Affiliations:
1external, ou_persistent22              
2Schwille, Petra / Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565169              

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Free keywords: TUG-OF-WAR; INTRACELLULAR-TRANSPORT; ORGANELLE TRANSPORT; MOLECULAR MOTORS; MYOSIN-V; PROTEINS; MOVEMENT; DYNAMICS; MICROTUBULES; TRACKINGScience & Technology - Other Topics; molecular motors; lipid bilayers; transport efficiency; motor-cargo coupling; streptavidin-binding peptide;
 Abstract: In eukaryotic cells, membranous vesicles and organelles are transported by ensembles of motor proteins. These motors, such as kinesin-1, have been well characterized in vitro as single molecules or as ensembles rigidly attached to nonbiological substrates. However, the collective transport by membrane-anchored motors, that is, motors attached to a fluid lipid bilayer, is poorly understood. Here, we investigate the influence of motors' anchorage to a lipid bilayer on the collective transport characteristics. We reconstituted "membrane-anchored" gliding motility assays using truncated kinesin-1 motors with a streptavidin-binding peptide tag that can attach to streptavidin-loaded, supported lipid bilayers. We found that the diffusing kinesin-1 motors propelled the microtubules in the presence of ATP. Notably, we found the gliding velocity of the microtubules to be strongly dependent on the number of motors and their diffusivity in the lipid bilayer. The microtubule gliding velocity increased with increasing motor density and membrane viscosity, reaching up to the stepping velocity of single motors. This finding is in contrast to conventional gliding motility assays where the density of surface-immobilized kinesin-1 motors does not influence the microtubule velocity over a wide range. We reason that the transport efficiency of membrane-anchored motors is reduced because of their slippage in the lipid bilayer, an effect that we directly observed using single-molecule fluorescence microscopy. Our results illustrate the importance of motor-cargo coupling, which potentially provides cells with an additional means of regulating the efficiency of cargo transport.

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Language(s): eng - English
 Dates: 2016-11-012016-11
 Publication Status: Issued
 Pages: 9
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: 000388970100008
DOI: 10.1073/pnas.1611398113
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Title: Proceedings of the National Academy of Sciences of the United States of America
  Other : Proc. Acad. Sci. USA
  Other : Proc. Acad. Sci. U.S.A.
  Abbreviation : PNAS
Source Genre: Journal
 Creator(s):
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Publ. Info: Washington, D.C. : National Academy of Sciences
Pages: - Volume / Issue: 113 (46) Sequence Number: - Start / End Page: E7185 - E7193 Identifier: ISSN: 0027-8424
CoNE: https://pure.mpg.de/cone/journals/resource/954925427230