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Query: UMLS:C0027960 (
mole
)
21,279
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Low concentrations of methanol, 2-propanol and ethylene glycol increase the asymmetry of the flagellar waveforms ad the turning rate of both live sperm and potentially symmetrical sperm reactivated with 1 mM-MgATP2-, while at the same time causing a decrease in the heat frequency. Similar effects are observed if the solvents are added to preparations of potentially symmetrical sperm reactivated in the presence of 1 mM free Ca2+, or to potentially asymmetrical sperm reactivated without added Ca2+, A second group of solvents, N,N-dimethylformamide, formamide and p-dioxane, also decrease the flagellar beat frequency, but have the opposite effect on symmetry, reducing the asymmetry of the waveforms and the turning rate of potentially symmetrical sperm reactivated in the presence of 1 mM free Ca2+. These effects of solvents are all reversible within about 5 min after initial exposure to solvent. Higher concentrations of methanol and 2-propanol (above approximately 5 and 0.8
mole
%, respectively) induce quiescence in potentially asymmetrical sperm reactivated with concentrations of MgATP2- ranging from 10 microM to 1 mM. The quiescent flagella initially assume a bent form very similar to that seen in Ca2+-induced quiescence, and show a subsequent time-dependent distortion of the initial bent from with eventual disintegration and splitting off of bundles of microtubules. Dimethylformamide, formamide and dioxane have almost no effect on the intrinsic asymmetry of potentially asymmetrical sperm reactivated in the absence of added Ca2+, but addition of these solvents to potentially asymmetrical sperm that have been induced to become quiescent by addition of 0.1 mM free Ca2+ causes the sperm to resume swimming with flagellar waveforms that are substantially more symmetrical that those of the starting preparation before the addition of Ca2+. Mild digestion with trypsin of reactivated sperm that have been induced either to beat asymmetrically or to become quiescent by addition of methanol causes a gradual appearance of symmetrical flagellar beating, as in the case of Ca2+-induced quiescence. The flagellar beat frequency, however, remains low, at about 20 Hz. The results suggest that the solvents either mimic or block the action of CA2+ by interaction with a Ca2+-dependent regulatory protein, and may also induce alteration in the rate constants of
dynein ATPase
.
...
PMID:Effects of organic solvents on flagellar asymmetry and quiescence in sea urchin sperm. 707 22
The binding of nucleoside triphosphates and related ligands to
dynein ATPase
from sea urchin sperm flagella has been studied by equilibrium partition analysis in an aqueous biphasic system containing dextran and poly(ethylene glycol). The stoichiometry of binding and the corresponding stepwise binding constants are obtained from direct binding isotherms fitted to the primary data. The results suggest that dynein possesses four different binding sites for nucleoside triphosphates per
mole
of heavy chain. The stepwise binding constants for MgATP range from approximately 10(4) M-1 to approximately 10(5) M-1. The isolated alpha and beta heavy chains have binding parameters similar to intact dynein. The amount of ADP bound normally is approximately 75% that of ATP, both for the intact dynein and for the separated heavy chains, although full saturation is achieved at high nucleotide concentrations. In the presence of the ATPase inhibitor vanadate, ADP binds with affinities similar to those of ATP, with binding constants close to those of ATP in the absence of vanadate. No appreciable binding of AMP or EDTA/ATP is observed. The substitution of Ca2+ or Fe3+ for Mg2+ does not significantly alter the amount of ATP bound; however, CaATP is bound with a somewhat lower affinity. Scatchard and Hill plots of the binding data and the calculated site-binding constants suggest that ATP and ADP bind in a weakly cooperative manner. These results suggest that the multiple binding of nucleotide to dynein heavy chains occurs at physiological concentrations, putatively at the four binding sites predicted earlier on the basis of their amino acid sequences. The data are consistent with a model in which, in addition to a single catalytic site, nucleotide binding occurs at additional noncatalytic sites that represent an as yet unknown functional aspect of dynein.
...
PMID:Phase partition analysis of nucleotide binding to axonemal dynein. 870 26