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Biochemical and Biophysical Research... Apr 1978
Topics: Adenosine Triphosphatases; Alkaline Phosphatase; Animals; Dyneins; Male; Mitosis; Myosins; Sea Urchins; Sperm Motility; Tetrahymena pyriformis; Vanadium
PubMed: 149544
DOI: 10.1016/0006-291x(78)91279-2 -
Journal of Biochemistry Aug 19791. Dynein was extracted with 0.5 M KCl from Tetrahymena axonemes. SDS-gel electrophoresis of the extract indicated that about 50% of the extracted protein had a...
1. Dynein was extracted with 0.5 M KCl from Tetrahymena axonemes. SDS-gel electrophoresis of the extract indicated that about 50% of the extracted protein had a molecular weight of about 3.5 X 10(5), and that 90% of the proteins with this weight had been extracted. 2. The ATPase [EC 3.6.1.3] reaction of the KCl-extracted dynein fraction was enhanced by 60-80% by addition of the outer doublet fraction. It showed an initial burst of Pi liberation of about 1 mol per mol of proteins with a molecular weight of 3.5 X 10(5). 3. We examined the interaction of the dynein-tubulin system from Tetrahymena cilia with ten ATP analogs [2'-dATP, 3'-dATP, epsilonATP, FTP, 8-NH(CH3)-ATP, 8,3'-S-cyclo-ATP, 8-Br-ATP, 8-OCH3-ATP, 8-SCH3-ATP, and AMPPNP]. Among them, 2'-dATP and 3'-dATP were good substrates for dynein ATPase, as they induced the dissociation of dynein arms from the B-tubule of outer doublets, the sliding movement between outer doublets, and the bending movement of axonemes. The other analogs did not induce the dissociation or the sliding movement. 4. Among the ATP analogs tested, only 2'-dATP and 3'-dATP induced the reorientation of cilia on the Triton model of Tetrahymena; the reorientation rates were smaller than that induced by ATP.
Topics: Adenosine Triphosphatases; Adenosine Triphosphate; Animals; Dyneins; Kinetics; Molecular Weight; Substrate Specificity; Tetrahymena pyriformis
PubMed: 158011
DOI: 10.1093/oxfordjournals.jbchem.a132540 -
Nature Feb 2003Dynein ATPases are microtubule motors that are critical to diverse processes such as vesicle transport and the beating of sperm tails; however, their mechanism of force...
Dynein ATPases are microtubule motors that are critical to diverse processes such as vesicle transport and the beating of sperm tails; however, their mechanism of force generation is unknown. Each dynein comprises a head, from which a stalk and a stem emerge. Here we use electron microscopy and image processing to reveal new structural details of dynein c, an isoform from Chlamydomonas reinhardtii flagella, at the start and end of its power stroke. Both stem and stalk are flexible, and the stem connects to the head by means of a linker approximately 10 nm long that we propose lies across the head. With both ADP and vanadate bound, the stem and stalk emerge from the head 10 nm apart. However, without nucleotide they emerge much closer together owing to a change in linker orientation, and the coiled-coil stalk becomes stiffer. The net result is a shortening of the molecule coupled to an approximately 15-nm displacement of the tip of the stalk. These changes indicate a mechanism for the dynein power stroke.
Topics: Adenosine Diphosphate; Animals; Chlamydomonas reinhardtii; Dyneins; Flagella; Microscopy, Electron; Movement; Protein Isoforms; Protein Structure, Tertiary; Structure-Activity Relationship; Vanadates
PubMed: 12610617
DOI: 10.1038/nature01377 -
Seikagaku. the Journal of Japanese... Oct 1988
Review
Topics: Adenosine Triphosphatases; Animals; Cilia; Dyneins; Flagella; Molecular Weight; Protein Conformation
PubMed: 2977787
DOI: No ID Found -
Plant Physiology Jun 2022Dynein motor proteins, often considered to be missing in land plants, are found in plants that reproduce with flagellated sperm.
Dynein motor proteins, often considered to be missing in land plants, are found in plants that reproduce with flagellated sperm.
Topics: Dyneins; Embryophyta; Phylogeny
PubMed: 35385107
DOI: 10.1093/plphys/kiac151 -
Proceedings of the National Academy of... Nov 1979Dynein isolated from Chlamydomonas flagellar axonemes binds to microtubules assembled in vitro from 6S brain tubulin dimers. The dynein arms bind periodically along the...
Dynein isolated from Chlamydomonas flagellar axonemes binds to microtubules assembled in vitro from 6S brain tubulin dimers. The dynein arms bind periodically along the length of the microtubules with a center-to-center spacing of 24 nm, equal to the periodicity of dynein arms on intact axonemes. The arms project from the in vitro assembled microtubules at an angle of approximately 55 degrees, thereby defining microtubule polarity. Dynein cosediments with microtubules through a sucrose gradient, as demonstrated by electron microscopy, gel electrophoresis, and ATPase analysis. In addition, dynein induces crossbridging between adjacent microtubules. Darkfield microscopy reveals that microtubules containing dynein are aggregated into large bundles; electron microscopy indicates that microtubules of the same polarity are crossbridged by a regular array of arms. Viewed by darkfield microscopy, addition of ATP to crossbridged microtubules causes their disaggregation; electron microscopy shows that the majority of these microtubules are no longer crossbridged. These observations are applicable to the determination of microtubule polarity and directionality of microtubule assembly in situ and suggest a role for dynein in cytoplasmic microtubule-based cellular movements.
Topics: Adenosine Triphosphatases; Animals; Brain; Carrier Proteins; Chlamydomonas; Cytoplasm; Dyneins; Flagella; Macromolecular Substances; Microtubules; Tubulin
PubMed: 160555
DOI: 10.1073/pnas.76.11.5759 -
TheScientificWorldJournal May 2010Unlike our understanding of the other two classes of cytoskeletal motor proteins, the myosins and kinesins, we have only recently begun to comprehend the molecular... (Review)
Review
Unlike our understanding of the other two classes of cytoskeletal motor proteins, the myosins and kinesins, we have only recently begun to comprehend the molecular mechanism for how dynein produces force and movement. The slow progress has been attributed, in part, to the enormous size of the dynein force-producing head, but also to the complex interplay between its structural components, each of which has a unique role in regulating dynein motor activity. The integrated and highly coordinated mechanism by which these structures work together in powering the dynein machinery is discussed in this review.
Topics: Dyneins; Protein Conformation
PubMed: 20454766
DOI: 10.1100/tsw.2010.76 -
Human Reproduction (Oxford, England) Aug 1994We tested the effects of adenosine and 2-deoxyadenosine on the activation of human spermatozoa. In the asthenozoospermic group of patients adenosine produces an increase...
We tested the effects of adenosine and 2-deoxyadenosine on the activation of human spermatozoa. In the asthenozoospermic group of patients adenosine produces an increase in sperm motility from 33.3 +/- 2.1% to 42.1 +/- 3.4%, progressive motility from 22.5 +/- 1.3% to 28.6 +/- 1.7% and forward progression rating from 2.1 +/- 0.2% to 2.8 +/- 0.1%. 2-Deoxyadenosine stimulated asthenozoospermic samples to a greater degree than adenosine. Sperm motility rose to 48.9 +/- 3.4%, progressive motility to 32.1 +/- 3.4% and forward progression rating to 3.0 +/- 0.1% following stimulation with 2-deoxy-adenosine. The kinetic parameters and basic characteristics of dynein ATPase were determined. The maximum activity of dynein ATPase, Vmax, was significantly different (P < 0.001) for asthenozoospermic and normozoospermic samples: 6.46 +/- 2.1 nmol Pi/mg/min and 16.99 +/- 3.7 nmol Pi/mg/min respectively. However, the enzyme affinity for ATP was not different. Stimulation of asthenozoospermic samples with adenosine and 2-deoxyadenosine caused an increase of Vmax (70-90% and 90-110% respectively) and no significant change in KM was observed. In order to block the nucleoside transporter and to eliminate the action of adenosine inside the cell, dipyridamole was used but the effects of adenosine were not neutralized. 5'-(N-ethylcarboxy-amido)-adenosine showed effects similar to those of adenosine, even when applied in 1 microM concentration. These results indicate that adenosine and its analogues stimulate sperm motility and activity of dynein ATPase, most probably via A2 receptors.
Topics: Adenosine; Adenosine-5'-(N-ethylcarboxamide); Deoxyadenosines; Dipyridamole; Dyneins; Humans; Infertility, Male; Kinetics; Male; Oligospermia; Sperm Motility; Spermatozoa
PubMed: 7989508
DOI: 10.1093/oxfordjournals.humrep.a138733 -
The Journal of Biological Chemistry Jul 2007According to the power stroke model of dynein deduced from electron microscopic and fluorescence resonance energy transfer studies, the power stroke and the recovery...
According to the power stroke model of dynein deduced from electron microscopic and fluorescence resonance energy transfer studies, the power stroke and the recovery stroke are expected to take place at the two isomerization steps of the ATPase cycle at the primary ATPase site. Here, we have conducted presteady-state kinetic analyses of these two isomerization steps with the single-headed motor domain of Dictyostelium cytoplasmic dynein by employing fluorescence resonance energy transfer to probe ATPase steps at the primary site and tail positions. Our results show that the recovery stroke at the first isomerization step proceeds quickly ( approximately 180 s(-1)), whereas the power stroke at the second isomerization step is very slow ( approximately 0.2 s(-1)) in the absence of microtubules, and that the presence of microtubules accelerates the second but not the first step. Moreover, a comparison of the microtubule-induced acceleration of the power stroke step and that of steady-state ATP hydrolysis implies the intriguing possibility that microtubules simultaneously accelerate the ATPase activity not only at the primary site but also at other site(s) in the motor domain.
Topics: Animals; Dictyostelium; Dyneins; Kinetics; Plasmids; Protozoan Proteins; Recombinant Fusion Proteins; Spectrometry, Fluorescence
PubMed: 17548361
DOI: 10.1074/jbc.M701914200 -
Nature Communications Dec 2017Cytoplasmic dynein is an enormous minus end-directed microtubule motor. Rather than existing as bare tracks, microtubules are bound by numerous microtubule-associated...
Cytoplasmic dynein is an enormous minus end-directed microtubule motor. Rather than existing as bare tracks, microtubules are bound by numerous microtubule-associated proteins (MAPs) that have the capacity to affect various cellular functions, including motor-mediated transport. One such MAP is She1, a dynein effector that polarizes dynein-mediated spindle movements in budding yeast. Here, we characterize the molecular basis by which She1 affects dynein, providing the first such insight into which a MAP can modulate motor motility. We find that She1 affects the ATPase rate, microtubule-binding affinity, and stepping behavior of dynein, and that microtubule binding by She1 is required for its effects on dynein motility. Moreover, we find that She1 directly contacts the microtubule-binding domain of dynein, and that their interaction is sensitive to the nucleotide-bound state of the motor. Our data support a model in which simultaneous interactions between the microtubule and dynein enables She1 to directly affect dynein motility.
Topics: Amino Acid Sequence; Dyneins; Microscopy, Fluorescence; Microtubule-Associated Proteins; Microtubules; Models, Biological; Models, Molecular; Molecular Motor Proteins; Myosin Heavy Chains; Myosin Type V; Protein Binding; Protein Domains; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins
PubMed: 29247176
DOI: 10.1038/s41467-017-02004-2