Subtilisin cleavage of actin inhibits in vitro sliding movement of actin filaments over myosin

Subtilisin cleaved actin was shown to retain several properties of intact actin including the binding of heavy meromyosin (HMM), the dissociation from HMM by ATP, and the activation of HMM ATPase activity. Similar Vmax but different Km values were obtained for acto- HMM ATPase with the cleaved and intact actins. The ATPase activity of HMM stimulated by copolymers of intact and cleaved actin showed a linear dependence on the fraction of intact actin in the copolymer. The most important difference between the intact and cleaved actin was observed in an in vitro motility assay for actin sliding movement over an HMM coated surface. Only 30% of the cleaved actin filaments appeared mobile in this assay and moreover, the velocity of the mobile filaments was approximately 30% that of intact actin filaments. These results suggest that the motility of actin filaments can be uncoupled from the activation of myosin ATPase activity and is dependent on the structural integrity of actin and perhaps, dynamic changes in the actin molecule.

• Publication year

  1990

• Venue

  Journal of Cell Biology

• Publication date

  1990-08-01

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1083/JCB.111.2.465PMID 2143196PMCID 2116201
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Propagation of Acto-S-1 ATPase reaction-coupled conformational change in actin along the filament.

  1989cited by this paper
• End-label fingerprintings show that the N- and C-termini of actin are in the contact site with gelsolin.

  1989cited by this paper
• Antibody against the amino terminus of alpha-actin inhibits actomyosin interactions in the presence of ATP.

  1989cited by this paper
• Molecular packing in profilin: actin crystals and its implications.

  1989cited by this paper
• Acanthamoeba actin and profilin can be cross-linked between glutamic acid 364 of actin and lysine 115 of profilin

  1989cited by this paper
• Calcium-triggered movement of regulated actin in vitro. A fluorescence microscopy study.

  1989cited by this paper
• Subtilisin-cleaved actin: polymerization and interaction with myosin subfragment 1.

  1989cited by this paper
• Polymerization of G-actin by myosin subfragment 1.

  1988cited by this paper
• Cross-linking of the skeletal myosin subfragment 1 heavy chain to the N-terminal actin segment of residues 40-113.

  1988cited by this paper
• Cation binding sites on actin: a structural relationship between antigenic epitopes and cation exchange.

  1988cited by this paper
• Modifying preselected sites on proteins: the stretch of residues 633-642 of the myosin heavy chain is part of the actin-binding site.

  1988cited by this paper
• The interaction of troponin-I with the N-terminal region of actin.

  1988cited by this paper
• Antibodies directed against N-terminal residues on actin do not block acto-myosin binding.

  1987cited by this paper
• Functional, chymotryptically split actin and its interaction with myosin subfragment 1.

  1987cited by this paper
• Anti-actin antibodies. An immunological approach to the myosin-actin and the tropomyosin-actin interfaces.

  1987cited by this paper
• Movement of myosin fragments in vitro: domains involved in force production.

  1987cited by this paper
• Myosin subfragment-1 is sufficient to move actin filaments in vitro

  1987cited by this paper
• Conformation of the ATP binding peptide in actin revealed by proton NMR spectroscopy.

  1987cited by this paper
• Spatial relationship between the nucleotide-binding site, Lys-61 and Cys-374 in actin and a conformational change induced by myosin subfragment-1 binding.

  1987cited by this paper
• Fluorescent actin filaments move on myosin fixed to a glass surface.

  1986cited by this paper
• The mechanism of muscle contraction.

  1986cited by this paper
• Myosin movement in vitro: a quantitative assay using oriented actin cables from Nitella.

  1986cited by this paper
• Angles of fluorescently labelled myosin heads and actin monomers in contracting and rigor stained muscle fiber.

  1984cited by this paper
• Chapter 3 – Macromolecular Assembly of Actin

  1983cited by this paper
• Identification of myosin-binding sites on the actin sequence.

  1982cited by this paper
• Effects of tryptic digestion on myosin subfragment 1 and its actin-activated adenosinetriphosphatase.

  1982cited by this paper
• Mechanism of action of troponin . tropomyosin. Inhibition of actomyosin ATPase activity without inhibition of myosin binding to actin.

  1981cited by this paper
• Structure of the actin–myosin interface

  1981cited by this paper
• Myosin minifilaments.

  1980cited by this paper
• On the origin of the contractile force in skeletal muscle.

  1979cited by this paper
• Rotational dynamics of spin-labeled F-actin in the sub-millisecond time range.

  1979cited by this paper
• Polarized fluorescence from epsilon-ADP incorporated into F-actin in a myosin-free single fiber: conformation of F-actin and changes induced in it by heavy meromyosin.

  1978cited by this paper
• Studies on the chymotryptic digestion of myosin. Effects of divalent cations on proteolytic susceptibility.

  1977influential reference
• Conformational change and cooperativity in actin filaments free of tropomyosin.

  1975cited by this paper
• Dynamic study of F-actin by quasielastic scattering of laser light.

  1971cited by this paper
• The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin.

  1971cited by this paper
• A mechanochemical mechanism for muscle contraction.

  1971cited by this paper
• Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

  1970cited by this paper

• Mechanisms of myosin II force generation. Insights from novel experimental techniques and approaches.

  2024cites this paper
• Photorhabdus luminescens TccC3 Toxin Targets the Dynamic Population of F-Actin and Impairs Cell Cortex Integrity

  2022cites this paper
• Myosin and gelsolin cooperate in actin filament severing and actomyosin motor activity

  2020cites this paper
• Actin.

  2020cites this paper
• Myosin-gelsolin cooperativity in actin filament severing and actomyosin motor activity

  2020cites this paper
• Uni-directional Propagation of Structural Changes in Actin Filaments

  2018cites this paper
• Chapter 17 Role of Dynamic and Cooperative Conformational Changes in Actin Filaments

  2016cites this paper
• Myopathy-causing Q147P TPM2 mutation shifts tropomyosin strands further towards the open position and increases the proportion of strong-binding cross-bridges during the ATPase cycle.

  2016cites this paper
• The Role of Structural Dynamics of Actin in Class-Specific Myosin Motility

  2015cites this paper
• Evidence for monomeric actin function in INO80 chromatin remodeling

  2013cites this paper
• Isolation and Sequencing of Actin 1 , Actin 2 and Tubulin 1 Genes Involved in Cytoskeleton Formation in

  2013cites this paper
• Isolation and Sequencing of Actin1, Actin2 and Tubulin1 Genes Involved in Cytoskeleton Formation in Phytophthora cinnamomi

  2013cites this paper
• Actin filaments as tension sensors.

  2012cites this paper
• 4.3 Structure and Dynamic States of Actin Filaments

  2012cites this paper
• Biochemical basis of human disease-causing actin mutations

  2011cites this paper
• Myosin binding surface on actin probed by hydroxyl radical footprinting and site-directed labels.

  2011cites this paper
• Remodeling of actin filaments by ADF/cofilin proteins

  2011cites this paper
• F-actin structure destabilization and DNase I binding loop: fluctuations mutational cross-linking and electron microscopy analysis of loop states and effects on F-actin.

  2010cites this paper
• Mutation of Actin Tyr-53 Alters the Conformations of the DNase I-binding Loop and the Nucleotide-binding Cleft*

  2010cites this paper
• Structural Polymorphism in F-actin

  2010cites this paper
• Screening of novel dominant negative mutant actins using glycine targeted scanning identifies G146V actin that cooperatively inhibits cofilin binding.

  2010cites this paper
• Slow skeletal muscle actin: confirmation and further insights

  2010cites this paper
• D-loop of Actin Differently Regulates the Motor Function of Myosins II and V*

  2009cites this paper
• Cell and molecular biology of the fastest myosins.

  2009cites this paper
• Molecular analysis of FER/actin complexes in prostate cancer

  2009cites this paper
• The in vitro motility assay parameters of actin filaments from Mytilus edulis exposed in vivo to copper ions.

  2009cites this paper
• Large-scale Models Reveal the Two-component Mechanics of Striated Muscle

  2008cites this paper
• Kinetic characterization of the function of myosin loop 4 in the actin-myosin interaction.

  2008cites this paper
• A Common Binding Site for Actin-Binding Proteins on the Actin Surface

  2007cites this paper
• Submolecular mechanisms underlying in vitro and in vivo effect of cardiac glycosides on contractile activity of myocardial myofibrils during heart failure

  2006cites this paper
• The DNase-I Binding Loop of Actin May Play a Role in the Regulation of Actin-Myosin Interaction by Tropomyosin/Troponin*

  2004cites this paper
• Actin-destabilizing factors disrupt filaments by means of a time reversal of polymerization.

  2004cites this paper
• Acrosomal actin: twists and turns of a versatile filament.

  2004cites this paper
• Actin-binding proteins--a unifying hypothesis.

  2004cites this paper
• The Regulation of Subtilisin-cleaved Actin by Tropomyosin/Troponin*

  2003cites this paper
• Crystal Structure of Monomeric Actin in the ATP State

  2003cites this paper
• Actin cross-linking and inhibition of the actomyosin motor.

  2002cites this paper
• Link between the enzymatic kinetics and mechanical behavior in an actomyosin motor.

  2001cites this paper
• Actin Depolymerizing Factor Stabilizes an Existing State of F-Actin and Can Change the Tilt of F-Actin Subunits

  2001cites this paper
• Probing the structure of F-actin: cross-links constrain atomic models and modify actin dynamics.

  2001cites this paper
• Divalent cations, nucleotides, and actin structure.

  2001cites this paper
• Actin allostery again?

  2001cites this paper
• F-actin retains a memory of angular order.

  2000cites this paper
• Proteolytic cleavage of actin within the DNase-I-binding loop changes the conformation of F-actin and its sensitivity to myosin binding.

  2000cites this paper
• Cross-linking constraints on F-actin structure.

  2000cites this paper
• Actin and the smooth muscle regulatory proteins: a structural perspective

  2000cites this paper
• Intermolecular dynamics and function in actin filaments.

  2000cites this paper
• Effect of intramolecular cross-linking between glutamine-41 and lysine-50 on actin structure and function

  2000cites this paper
• Conformational changes of contractile proteins and their role in muscle contraction.

  1999cites this paper
• Intrastrand cross-linked actin between Gln-41 and Cys-374. III. Inhibition of motion and force generation with myosin.

  1998cites this paper
• Cooperative rigor binding of myosin to actin is a function of F-actin structure.

  1997cites this paper
• Perturbations of functional interactions with myosin induce long-range allosteric and cooperative structural changes in actin.

  1997cites this paper
• Actin: From cell biology to atomic detail

  1997cites this paper
• The possible role of myosin A1 light chain in the weakening of actin-myosin interaction.

  1997cites this paper
• New angles on actin dynamics.

  1997cites this paper
• Myosin-induced changes in F-actin: fluorescence probing of subdomain 2 by dansyl ethylenediamine attached to Gln-41.

  1996cites this paper
• Phase transitions and the molecular mechanism of contraction.

  1996cites this paper
• Cooperativity in F-actin: binding of gelsolin at the barbed end affects structure and dynamics of the whole filament.

  1996cites this paper
• The structure of an open state of beta-actin at 2.65 A resolution.

  1996cites this paper
• [The mechanism of muscle contraction: along the path carved by Academician G.M Frank].

  1996cites this paper
• Intermolecular coupling between loop 38-52 and the C-terminus in actin filaments.

  1996cites this paper
• Purification and characterization of subtilisin cleaved actin lacking the segment of residues 43-47 in the DNase I binding loop.

  1995cites this paper
• Structural dynamics of F-actin: I. Changes in the C terminus.

  1995cites this paper
• A novel kDa form of subtilisin cleaved actin: structural and functional consequences of cleavage between Ser234 and Ser235

  1995cites this paper
• Muscle contraction mechanism: are alternative engines gathering steam?

  1995cites this paper
• Allostery, cooperativity, and different structural states in F-actin.

  1995cites this paper
• Immunochemical probing of the functional role of the 238-246 and 567-574 sequences of myosin heavy chain.

  1995cites this paper
• New insights into actin filament dynamics.

  1995cites this paper
• end5, end6, and end7: mutations that cause actin delocalization and block the internalization step of endocytosis in Saccharomyces cerevisiae.

  1995cites this paper
• The effects of phosphorylation of cardiac troponin-I on its interactions with actin and cardiac troponin-C.

  1995cites this paper
• Two identical hydrophobic clusters are present on the same actin monomer: interaction between one myosin subfragment‐1 and two actin monomers

  1995cites this paper
• Actin and the actomyosin interface: a review.

  1995cites this paper
• Dynamic properties of actin. Structural changes induced by beryllium fluoride.

  1994cites this paper
• A Model of the Leukocyte Migration Through Solid Tissue

  1994cites this paper
• Microsecond rotational dynamics of F-actin in ActoS1 filaments during ATP hydrolysis.

  1994cites this paper
• Orientation of actin filaments during motion in in vitro motility assay.

  1994cites this paper
• Cooperativity in F-actin: chemical modifications of actin monomers affect the functional interactions of myosin with unmodified monomers in the same actin filament.

  1993cites this paper
• Inhibition of actin filament movement by monoclonal antibodies against the motor domain of myosin

  1993cites this paper
• Motility assay: achievements and perspectives.

  1993cites this paper
• In vitro motility of proteolytically cleaved myosin subfragment 1 on a lysine-coated surface.

  1993cites this paper
• CHARACTERIZATION OF SKELETAL MUSCLE ACTIN LABELED WITH THE TRIPLET PROBE ERYTHROSIN‐5‐IODOACETAMIDE

  1993cites this paper
• In vitro movement of actin filaments over myosin: role of actin and related problems.

  1993cites this paper
• Actin molecular structure and function.

  1993cites this paper
• How are myosin fragments bound to nitrocellulose film?

  1993cites this paper
• Smooth and skeletal muscle actin are mechanically indistinguishable in the in vitro motility assay.

  1993cites this paper
• Myosin subfragment 1 inhibits dissociation of nucleotide and calcium from G-actin.

  1993cites this paper
• Mutations in β‐actin: influence on polymer formation and on interactions with myosin and profilin

  1993cites this paper
• Molecular genetics of actin function.

  1993cites this paper
• Enhanced stimulation of myosin subfragment 1 ATPase activity by addition of negatively charged residues to the yeast actin NH2 terminus.

  1993cites this paper
• Actin as the generator of tension during muscle contraction.

  1992cites this paper
• Cooperativity of thiol-modified myosin filaments. ATPase and motility assays of myosin function.

  1992cites this paper
• Contraction due to bimetallic, short-lived super-coiling in the helical, double stranded filaments of striated muscle.

  1992cites this paper
• Muscle contraction and in vitro actin? movement: role of

  1992cites this paper
• Muscle actin cleaved by proteinase K: its polymerization and in vitro motility.

  1992cites this paper
• Yeast actin filaments display ATP-dependent sliding movement over surfaces coated with rabbit muscle myosin.

  1992cites this paper
• Muscle contraction andin vitro movement: Role of actin?

  1992cites this paper
• Site-directed mutations of Dictyostelium actin: disruption of a negative charge cluster at the N terminus.

  1991cites this paper