DOI: 10.1111/j.1432-1033.1989.tb14826.x. The complex of actin and deoxyribonuclease I as a model system to study the interactions of nucleotides, cations and cytochalasin D with monomeric actin

The complex of actin and deoxyribonuclease I as a model system to study the interactions of nucleotides, cations and cytochalasin D with monomeric actin

10.1111/j.1432-1033.1989.tb14826.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

The stoichiometric actin – DNase‐I complex was used to study the actin – nucleotide and actin – divalentcation interactions and its ATPase activity in the presence of MgCl2 and cytochalasin D. Treatment of actin – DNase‐I complex with 1 mM EDTA results in almost complete restoration of its otherwise inhibited DNase I activity, although the complex does not dissociate, as verified by size‐exclusion chromatography. This effect is due to a loss of actin‐bound nucleotide but is prevented by the presence of 0.1 – 0.5 mM ATP, ADP and certain ATP analogues. In this case no increase in DNase I activity occurs, even in the presence of EDTA. At high salt concentrations and in the presence of Mg2+ (‘physiological conditions’) the association rate constants for ATP, ADP and ɛATP (1,N6‐ethenoadenosine 5′‐triphosphate) and the dissociation rate constant for ɛATP were determined. Both the on and off rates were found to be reduced by a factor about 10 when compared to uncomplexed actin. Thus the binding constant of ɛATP to actin is almost unaltered after complexing to DNase I (2.16 × 108 M−1). Titrating the increase in DNase I activity of the actin – DNase I complex against nucleotide concentration in the presence of EDTA, the association constant of ATP to the cation‐free form of actin – DNase I complex was found to be 5 × 103 M−1, which is many orders of magnitude lower than in the presence of divalent metal ions. The binding constant of Ca2+ to the high‐affinity metal‐binding site of actin was found not to be altered when complexed to DNase I, although the rate of Ca2+ release decreases by a factor of 8 after actin binding to DNase I. The rate of denaturation of nucleotide‐free and metal‐ion‐free actin – DNase I complex was found to be reduced by a factor of about 15. The ATPase activity of the complex is stimulated by addition of Mg2+ and even more effectively by cytochalasin D, proving that this drug is able to interact with monomeric actin.

Bibliography

POLZAR, B., NOWAK, E., GOODY, R. S., & MANNHERZ, H. G. (1989). The complex of actin and deoxyribonuclease I as a model system to study the interactions of nucleotides, cations and cytochalasin D with monomeric actin. European Journal of Biochemistry, 182(2), 267â275. Portico.

Authors 4

Bernhard POLZAR (first)
Ewa NOWAK (additional)
Roger S. GOODY (additional)
Hans Georg MANNHERZ (additional)

References 29 Referenced 22

10.1146/annurev.bi.55.070186.005011
10.1073/pnas.71.12.4742
10.1016/0014-5793(75)80412-1
10.1016/0092-8674(76)90203-8
10.1111/j.1432-1033.1980.tb04437.x
10.1016/S0021-9258(19)70681-4
10.1021/bi00263a009
10.1111/j.1432-1033.1975.tb02320.x
10.1016/0006-291X(86)90036-7
10.1085/jgp.33.4.349
10.1038/227680a0
10.1021/bi00769a001
10.1021/ja00752a042
10.1021/bi00653a015
10.1021/bi00789a009
10.1111/j.1432-1033.1975.tb02291.x
10.1016/0167-4838(85)90190-6
10.1111/j.1432-1033.1977.tb11385.x
10.1021/bi00405a060
10.1016/0014-5793(83)81059-X
10.1016/S0021-9258(19)86107-0
10.1016/S0021-9258(19)68895-2 / J. Biol. Chem. by Brenner S. L. (1981)
10.1111/j.1432-1033.1984.tb08018.x
10.1021/bi00423a016
10.1016/S0021-9258(18)88820-2
10.1002/j.1460-2075.1985.tb03900.x
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10.1083/jcb.84.2.455
10.1016/S0021-9258(18)66662-1 / J. Biol. Chem. by Goddette D. W. (1986)

Dates

Type	When
Created	20 years, 5 months ago (March 3, 2005, 10:12 p.m.)
Deposited	1 year, 9 months ago (Nov. 23, 2023, 5:58 p.m.)
Indexed	2 months, 3 weeks ago (June 10, 2025, 3:04 p.m.)
Issued	36 years, 3 months ago (June 1, 1989)
Published	36 years, 3 months ago (June 1, 1989)
Published Online	20 years, 5 months ago (March 3, 2005)
Published Print	36 years, 3 months ago (June 1, 1989)

Funders 0

None

BibTeX

@article{POLZAR_1989, title={The complex of actin and deoxyribonuclease I as a model system to study the interactions of nucleotides, cations and cytochalasin D with monomeric actin}, volume={182}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1989.tb14826.x}, DOI={10.1111/j.1432-1033.1989.tb14826.x}, number={2}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={POLZAR, Bernhard and NOWAK, Ewa and GOODY, Roger S. and MANNHERZ, Hans Georg}, year={1989}, month=jun, pages={267–275} }

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