Reaction mechanism of 21S dynein ATPase from sea urchin sperm. I. Kinetic properties in the steady state.

Reaction mechanism of 21S dynein ATPase from sea urchin sperm. I. Kinetic properties in the steady state. Research Abstract Details 

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Reaction mechanism of 21S dynein ATPase from sea urchin sperm. I. Kinetic properties in the steady state. Abstract Text:

S Terashita,T Kato,H Sato,

21S Dynein ATPase [EC 3.6.1.3] from axonemes of a Japanese sea urchin, Pseudocentrotus depressus, and its subunit fractions were studied to determine their kinetic properties in the steady state, using [gamma-32P]ATP at various concentrations, 5 mM divalent cations, and 20 mM imidazole at pH 7.0 and 0 degrees C. The following results were obtained. 1. 21S Dynein had a latent ATPase activity of about 0.63 mumol Pi/(mg . min) in 1 mM ATP, 100 mM KCl, 4 mM MgSO4, 0.5 mM EDTA, and 30 mM Tris-HCl at pH 8.0 and 25 degrees C. Its exposure to 0.1% Triton X-100 for 5 min at 25 degrees C induced an increase in the ATPase activity to about 3.75 mumol Pi/(mg . min) and treatment at 40 degrees C for 5 min also induced a similar activation. 2. The double-reciprocal plot for the ATPase activity of dynein activated by the treatment at 40 degrees C consisted of two straight lines, while that of nonactivated 21S dynein fitted a single straight line. 3. In low ionic strength solution, the Mg- and Mn-ATPase of 21S dynein showed substrate inhibition at ATP concentrations above 0.1 mM; the inhibition decreased with increasing ionic strength. Ca- and Sr-ATPase showed no substrate inhibition. 4. Both the Vmax and Km values of dynein ATPase decreased reversibly upon addition of about 40% (v/v) glycerol. In the presence of glycerol, the dynein ATPase showed an initial burst of Pi liberation. The apparent Pi-burst size was 1.0 mol/(10(6) g protein) and the true size was calculated to be 1.6 mol/1,250 K after correcting for the effect of Pi liberation in the steady state and the purity of our preparation. 5. One of the subunit fractions of 21S dynein which was obtained by the method of Tang et al. showed substrate inhibition and an initial burst of Pi liberation of 1.4 mol/(10(6) g protein) in the presence of 54% (v/v) glycerol.

Reaction mechanism of 21S dynein ATPase from sea urchin sperm. I. Kinetic properties in the steady state. Publishing Authors By Initials

S Terashita,T Kato,H Sato,

For similar urogenital system: genitalia: germ cells: spermatozoa research abstracts see: urogenital system: genitalia: germ cells: spermatozoa research

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Reaction mechanism of 21S dynein ATPase from sea urchin sperm. I. Kinetic properties in the steady state. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biochemistry

VOLUME: 93

Page Numbers: 1567-74

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Jun

YEAR: 1983

Reaction mechanism of 21S dynein ATPase from sea urchin sperm. I. Kinetic properties in the steady state. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 376600

Reaction mechanism of 21S dynein ATPase from sea urchin sperm. I. Kinetic properties in the steady state. Keywords Mesh Terms:

KEYWORDS: Spermatozoa

MESH TERMS: enzymology

Chemical & Substance for Abstract: Reaction mechanism of 21S dynein ATPase from sea urchin sperm. I. Kinetic properties in the steady state. Information

Substance Name: Dynein ATPase

Registry Number: EC 3.6.4.2

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Country: JAPAN

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MEDLINETA: J Biochem

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