Studies on the amino groups of myosin ATPase

Mühlrád, András; Srivastava, Sudhir; Hollósi, G; Wikman‐Coffelt, Joan

doi:10.1016/0003-9861(81)90285-x

Cited by 11 publications

(5 citation statements)

References 22 publications

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“…K + EDTA-ATPases are 17 or 26% of control, Ca 2+ -ATPases are 110 or 95%, and Mg 2+ -ATPases are 800 or 426%. The changes in the different ATPase activities upon TNP modification of S1 or βS1 shows 70−80% of the label resides on Lys83 as shown previously ( , ).…”

Section: Methodssupporting

confidence: 83%

“…β-Cardiac myosin was prepared from bovine heart ventriculum by combination of the protocols of Tada et al (41) and Muhlrad et al (7) with several modifications. Myosin was extracted from minced, washed ventriculum for 10 min at 4 °C with "Guba-Straub" solution containing 0.3 M KCl, 1 mM MgCl 2 , 10 mM sodium pyrophosphate, 1 mM DTT, and 0.15 M potassium phosphate buffer, pH 6.5.…”

Section: Methodsmentioning

confidence: 99%

“…Chemical Modification of Proteins. Skeletal or cardiac S1 was trinitrophenylated according to published protocols ( , ). A 2.2-fold molar excess of TNBS was added to 60−80 μM S1 in 30 mM KCl and 0.2 mM PMSF, 100 mM Tris-HCl buffer, pH 7.8. βS1 was trinitrophenylated in the same condition except the KCl concentration was 0.5 M. After incubation at 25 °C for 10 min, the reaction was terminated by addition of 2 mM DTT.…”

Section: Methodsmentioning

confidence: 99%

“…Cardiac myosin, the motor protein functioning in the heart as the chemomechanical energy transducer (2), associates with actin to transduce chemical energy in ATP to mechanical work. The globular head portion of cardiac myosin, subfragment 1 (S1), 1,2 shares sequence (79% identity), the ATP hydrolysis scheme (3,4), the characteristics of an ATP sensitive tryptophan (3)(4)(5), a highly reactive thiol (6) and lysine residue (7), proteolytic digestion patterns (8), and other physical properties with skeletal myosin S1, suggesting their comparison in the investigation of the molecular mechanism of energy transduction (9). The skeletal and cardiac S1 analogy offers many points for comparison of which we involve two.…”

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confidence: 99%

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Structural Characterization of β-Cardiac Myosin Subfragment 1 in Solution

et al. 2001

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beta-cardiac myosin subfragment 1 (betaS1) tertiary structure and dynamics were characterized with proteolytic digestion, nucleotide analogue trapping kinetics, and intrinsic fluorescence changes accompanying nucleotide binding. Proteolysis of betaS1 produces the 25, 50, and 20 kDa fragments and a new cut within the 50-kDa fragment at Arg369. F-actin inhibits cleavage of the 50-kDa fragment and fails to inhibit cleavage at the 50/20 kDa junction, suggesting betaS1 presents an actoS1 conformation fundamentally different from skeletal S1. Time-dependent changes in Mg(2+)-ATPase accompanying proteolysis identifies cleavage points that lie within the energy transduction pathway. The nucleotide analogue trapping kinetics reveal the presence of a reversible weakly actin attached state. Comparison of nucleotide analogue induced betaS1 structures with the transient structures occurring during ATPase indicates analogue induced and transient structures are in a one-to-one correspondence. Tryptophan fluorescence enhancement accompanies the binding or trapping of nucleotide or nucleotide analogues. Isolation of Trp508 fluorescence shows it is an ATP-sensitive tryptophan and that its vicinity changes conformation sequentially with the transient intermediates accompanying ATPase. These studies elucidate energy transduction and suggest how mutations of betaS1 implicated in disease might undermine function, stability, or efficiency.

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Section: Methodssupporting

confidence: 83%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Structural Characterization of β-Cardiac Myosin Subfragment 1 in Solution

et al. 2001

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“…Cardiac S1 also possesses a reactive lysyl residue (RLR) and an arginine residue in the equivalent positions to Lys84 and Arg723 in chicken skeletal myosin; however, Tyr85 is replaced by Phe. Cardiac RLR reaction rates are 2-3-fold less than that of skeletal RLR but still anomalously higher than the other exposed lysines (57). This observation implies a role for Tyr85 in the anomalous behavior of RLR, possibly as an additional proton acceptor from Lys84.…”

Section: Discussionmentioning

confidence: 84%

Trinitrophenylated Reactive Lysine Residue in Myosin Detects Lever Arm Movement during the Consecutive Steps of ATP Hydrolysis

Ajtai¹,

Ym²,

Park³

et al. 1999

Biochemistry

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Trinitrophenylation of the reactive lysine (Lys84) in skeletal myosin subfragment 1 (S1) introduces a chiral probe (TNP) into an interface of the catalytic and lever arm domains of S1 [Muhlrad (1977) Biochim. Biophys. Acta 493, 154-166]. Characteristics of the TNP absorption and circular dichroism (CD) spectra in TNP-modified S1 (TNP-Lys84-S1), and the Lys84 trinitrophenylation rate in native S1, indicate a one-to-one correspondence between ATPase transients and trapped phosphate analogues. Phosphate analogue-induced structures of TNP-Lys84-S1 were modeled using the crystallographic coordinates of S1 [Rayment et al. (1993) Science 261, 50-58] with swivels at Gly699 and Gly710 to approximate conformational changes during ATPase. The CD and absorption spectral characteristics of the model structures were compared to those observed for analogue-induced structures. The model calculations, first tested on a trinitrophenylated hexapeptide with known structure, were applied to TNP-Lys84-S1. They showed that ATP binding initiates swiveling at Gly699 and that swiveling at both Gly710 and Gly699 accompanied ATP splitting just prior to product release. The computed lever arm trajectory during ATPase suggests (i) a plausible mechanism for the nucleotide-induced inhibition of Lys84 trinitrophenylation, and (ii) trinitrophenylation-induced changes in S1 Mg2+- and K+-EDTA ATPase are from collision of the lever arm with TNP at Lys84. TNP is a site-specific structural perturbant of S1 and a chiral reporter group for the effect of Lys84 modification on dynamic S1 structure. As such, TNP-Lys84-S1 is equivalent to a genetically engineered mutant with intrinsic sensitivity to structure local to the modified residue.

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Conformational Changes and Molecular Dynamics of Myosin

Gergely

Seidel

1983

Comprehensive Physiology

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The sections in this article are: Myosin Structure Subunits Actin Interaction Site Myosin Interaction with ATP and its Analogues Conformational Changes Ultraviolet Spectral Changes Electron Paramagnetic Resonance Spectra Relation to ATPase Mechanism Rod Conformational Changes Chemical Modification Cysteine Residue Modification Lysine Modification Two‐Headed Nature of Myosin Are the Two Myosin Heads Identical? Interactions of the Two Myosin Heads Molecular Mechanism of Cross‐Bridge Cycle Segmental Flexibility of Myosin Measurement of Correlation Time Local Fluctuations in Myosin Structure Organized Systems Motion of Myosin Heads Orientation of Myosin Heads Force Generation and Elasticity Rotating‐Head Models Helix‐Coil Melting Model Actin Motions X‐Ray Studies Correlation Functions

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Studies on the amino groups of myosin ATPase

Cited by 11 publications

References 22 publications

Structural Characterization of β-Cardiac Myosin Subfragment 1 in Solution

Structural Characterization of β-Cardiac Myosin Subfragment 1 in Solution

Trinitrophenylated Reactive Lysine Residue in Myosin Detects Lever Arm Movement during the Consecutive Steps of ATP Hydrolysis

Conformational Changes and Molecular Dynamics of Myosin

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