Enhanced stimulation of myosin subfragment 1 ATPase activity by addition of negatively charged residues to the yeast actin NH2 terminus

R Kimberley Cook; Douglas Root; Carl Miller; Emil Reisler; Peter A Rubenstein

doi:10.1016/S0021-9258(18)53791-1

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Enhanced stimulation of myosin subfragment 1 ATPase activity by addition of negatively charged residues to the yeast actin NH2 terminus

Journal article

Open access

Peer reviewed

Enhanced stimulation of myosin subfragment 1 ATPase activity by addition of negatively charged residues to the yeast actin NH2 terminus

R Kimberley Cook, Douglas Root, Carl Miller, Emil Reisler and Peter A Rubenstein

The Journal of biological chemistry, Vol.268(4), pp.2410-2415

02/05/1993

DOI: 10.1016/S0021-9258(18)53791-1

PMID: 8428914

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Abstract

We examined the effects of yeast actin NH2-terminal mutations on actomyosin interactions and the function of actin in vivo through measurements of actin-activated ATPase activity, cosedimentation with rabbit muscle myosin subfragment 1 (S-1), in vitro motility, and invertase secretion assays. As reported earlier (Cook, R. K., Blake, W., and Rubenstein, P. A. (1992) J. Biol. Chem. 267, 9430-9436), elimination of NH2-terminal acidic residues from yeast actin results in an increased actin bundling, decreased actin-activated S-1 ATPase, and complete inhibition of actin filament sliding over myosin. Here we show that the addition of 2 new acidic residues to the NH2 terminus of yeast actin increased the Vmax value and the catalytic efficiency of the actin-activated ATPase activity of S-1. However, the binding of actin to S-1 in the presence of ATP and the velocities of actin sliding over myosin in the in vitro motility assays were not affected by this mutation. Thus, the number of actin NH2-terminal negative charges is important for actin activation of myosin S-1 ATPase activity, while only a minimum number of acidic residues is required for actin sliding over myosin in vitro. The number of actin NH2-terminal negative charges therefore appears to determine the efficiency with which the energy from ATP hydrolysis is converted to filament sliding.

Amino Acid Sequence

Rabbits

Mutagenesis, Site-Directed

Myosin Subfragments - metabolism

Actins - metabolism

Oligodeoxyribonucleotides - chemistry

Molecular Sequence Data

Structure-Activity Relationship

Saccharomyces cerevisiae - chemistry

Macromolecular Substances

Animals

Muscle Contraction

Base Sequence

Actins - chemistry

Myosins - metabolism

Enzyme Activation

Kinetics

In Vitro Techniques

Fungal Proteins - metabolism

Glycoside Hydrolases - secretion

beta-Fructofuranosidase

Details

Title: Subtitle: Enhanced stimulation of myosin subfragment 1 ATPase activity by addition of negatively charged residues to the yeast actin NH2 terminus
Creators: R Kimberley Cook - Department of Biochemistry, College of Medicine, University of Iowa, Iowa City 52242
Douglas Root
Carl Miller
Emil Reisler
Peter A Rubenstein
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.268(4), pp.2410-2415
DOI: 10.1016/S0021-9258(18)53791-1
PMID: 8428914
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: AR 22031 / NIAMS NIH HHS GM-33689 / NIGMS NIH HHS R01 AR022031 / NIAMS NIH HHS
Language: English
Date published: 02/05/1993
Academic Unit: Stead Family Department of Pediatrics; Biochemistry and Molecular Biology; Internal Medicine
Record Identifier: 9984024413202771

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