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Effects of N-Ethylmaleimide on Dynein Adenosinetriphosphatase Activity and Its Recombining Ability with Outer Fibers
Abstract
The effects of N-ethylmaleimide (NEM) on ciliary ATPase (dyneins) [EC 3.6.1.3] of Tetrahymena pyriformis were investigated. Like muscle myosin ATPase, 30 S dynein ATPase was activated biphasically by NEM, while 14 S dynein ATPase was simply inhibited. The maximal activation of 30 S dynein ATPase was obtained by preincubation with NEM at a concentration of 50 μM (0.23 μmole NEM/mg protein) and the magnitude of activation was 2.5- to 3-fold. The changes in the characteristics of 30 S dynein ATPase caused by modification were studied in comparison with those of myosin ATPase. It was confirmed that 30 S dynein ATPase was a myosin-like enzyme, though a few differences were found. From recombination tests of modified 30 S dynein with outer fibers, it was shown that at least three types of -SH groups are present in 30 S dynein molecules; -SH groups responsible for recombination with outer fibers, for enhancement of the enzyme activity by their blockage, and for catalytic activity.
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“While NEM inhibits force generation by dynein (Mitchell and Warner, 1981; Cosson and Gibbons, 1978), as well as dynein’s ability to bind to microtubules (Shimizu and Kimura, 1974)”
From Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility (1985) · cites this paper as Shimizu and Kimura, 1974
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Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility
VALE, R., REESE, T., & SHEETZ, M. (1985) · Cell · doi:10.1016/s0092-8674(85)80099-4 · Cited by 1,993
Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro
Vale, R. D., Schnapp, B. J., Mitchison, T. J., Steuer, E. R., Reese, T. S., & Sheetz, M. P. (1985) · Cell · doi:10.1016/0092-8674(85)90234-x · Cited by 462
Characterization of the microtubule-activated ATPase of brain cytoplasmic dynein (MAP 1C).
Shpetner, H. S., Paschal, B. M., & Vallee, R. B. (1988) · The Journal of Cell Biology · doi:10.1083/jcb.107.3.1001 · Cited by 140
Chemical subdomains within the kinetochore domain of isolated CHO mitotic chromosomes.
Wordeman, L. G., Steuer, E. R., Sheetz, M. P., & Mitchison, T. J. (1991) · The Journal of Cell Biology · doi:10.1083/jcb.114.2.285 · Cited by 104
Design and regulation of the AAA+ microtubule motor dynein
Sakato, M., & King, S. M. (2003) · Journal of Structural Biology · doi:10.1016/j.jsb.2003.09.026 · Cited by 100
Metadata
- Authors
- Shimizu, T., & KIMURA, I.
- Affiliations
- The University of Tokyo (JP)
- Venue
- The Journal of Biochemistry
- Publisher
- Oxford University Press (OUP)
- Published
- 1974-11
- Volume
- 76
- Issue
- 5
- Pages
- 1001-8
- ISSN
- 0021-924X (ISSN-L)
- 1756-2651
- OpenAlex
- W884304900
- Cited by
- 41
- References
- 0
- Citations by year
- 2023: 1 · 2017: 1 · 2014: 1 · 2012: 1
- FWCI
- 2.03
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- The Journal of Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic and Kidney Cyst Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › ATP Synthase and ATPases Research
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Cystic Fibrosis Research Advances
- Keywords
- N-ethylmaleimide, dynein ATPase, Tetrahymena pyriformis, enzyme activation, sulfhydryl groups, catalytic activity
- MeSH terms
- Adenosine Triphosphatases, Cilia, Enzyme Activation, Ethylmaleimide, Hydrogen-Ion Concentration, Potassium Chloride, Temperature, Tetrahymena, Trichloroacetic Acid
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