Batters, Christopher, Woodall, Katy A, Toseland, Christopher P., Hundschell, Christian, Veigel, Claudia (2012) Cloning, expression, and characterization of a novel molecular motor, Leishmania myosin-XXI. Journal of Biological Chemistry, 287 (33). pp. 27556-27566. ISSN 0021-9258. (doi:10.1074/jbc.M112.381301) (Access to this publication is currently restricted. You may be able to access a copy if URLs are provided) (KAR id:47851)
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Official URL: http://dx.doi.org/10.1074/jbc.M112.381301 |
Abstract
The genome of the Leishmania parasite contains two classes of myosin. Myosin-XXI, seemingly the only myosin isoform expressed in the protozoan parasite, has been detected in both the promastigote and amastigote stages of the Leishmania life cycle. It has been suggested to perform a variety of functions, including roles in membrane anchorage, but also long-range directed movements of cargo. However, nothing is known about the biochemical or mechanical properties of this motor. Here we designed and expressed various myosin-XXI constructs using a baculovirus expression system. Both full-length (amino acids 1-1051) and minimal motor domain constructs (amino acids 1-800) featured actin-activated ATPase activity. Myosin-XXI was soluble when expressed either with or without calmodulin. In the presence of calcium (pCa 4.1) the full-length motor could bind a single calmodulin at its neck domain (probably amino acids 809-823). Calmodulin binding was required for motility but not for ATPase activity. Once bound, calmodulin remained stably attached independent of calcium concentration (pCa 3-7). In gliding filament assays, myosin-XXI moved actin filaments at ?15 nm/s, insensitive to both salt (25-1000 mm KCl) and calcium concentrations (pCa 3-7). Calmodulin binding to the neck domain might be involved in regulating the motility of the myosin-XXI motor for its various cellular functions in the different stages of the Leishmania parasite life cycle.
Item Type: | Article |
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DOI/Identification number: | 10.1074/jbc.M112.381301 |
Uncontrolled keywords: | Cytoskeleton; Enzyme Kinetics; Leishmania; Molecular Motors; Myosin |
Subjects: | Q Science |
Divisions: | Divisions > Division of Natural Sciences > Biosciences |
Depositing User: | Chris Toseland |
Date Deposited: | 07 Apr 2015 11:55 UTC |
Last Modified: | 16 Nov 2021 10:19 UTC |
Resource URI: | https://kar.kent.ac.uk/id/eprint/47851 (The current URI for this page, for reference purposes) |
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