PMID: 2500340Jun 15, 1989Paper

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
B PolzarHans Georg Mannherz

Abstract

The stoichiometric actin--DNase-I complex was used to study the actin--nucleotide and actin--divalent-cation 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 epsilon ATP (1,N6-ethenoadenosine 5'-triphosphate) and the dissociation rate constant for epsilon ATP were determined. Both the on and off rates were found to be reduced by a factor of about 10 when compared to uncomplexed actin. Thus the binding constant of epsilon ATP to actin is almost unaltered after complexing to DNase I (2.16 x 10(8) M-1). Titrating the increase in DNase I activity of the actin--DNase I complex against nucleotide concentration in the presence of EDTA, ...Continue Reading

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Citations

Jan 1, 1994·Cell Motility and the Cytoskeleton·J M AnderslandM V Parthasarathy
May 1, 2007·Apoptosis : an International Journal on Programmed Cell Death·Dirk Eulitz, Hans Georg Mannherz
Nov 5, 1992·Journal of Immunological Methods·U E GibsonD V Sinicropi
Dec 1, 1992·Plant Physiology·J M AnderslandM V Parthasarathy
Oct 1, 1992·European Journal of Biochemistry·D R DrummondJ C Sparrow
Nov 1, 2008·The International Journal of Biochemistry & Cell Biology·Sebastian LudwigMarkus Napirei
Sep 26, 2003·Biophysical Journal·Dmitry S Kudryashov, Emil Reisler
Sep 6, 1990·Nature·W KabschK C Holmes
Jun 1, 1992·Journal of Muscle Research and Cell Motility·J E EstesL C Gershman
Nov 3, 2001·Biochimica Et Biophysica Acta·H Schüler

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