Involvement of the lipid and protein components of (Na+ + K+)-adenosine triphosphatase in the inhibitory action of alcohol

1. An experimental study was made on the adenosine triphosphatase action of crystalline myosin and actomyosin preparations under different conditions. 2. No enzymatic activity was found in the absence of salts. Activation was given by KCl and CaCl2, whereas MgCl2 in the presence of other ions inhibited. 3. The effect of pH is complex. In stabilizing buffers or at low temperature, there are two optima (pH 6.2 to 6.5 and pH 9.2) provided Ca is present. Without Ca only the acid optimum is found. The highest activities are reached in glycine buffer at pH 9.2 in the presence of Ca. 4. The study of the Mg-Ca antagonism revealed that the inhibition due to Mg is fully developed with Mg:Ca ratios less than 1, the inhibition usually exceeding 90 per cent. 5. It is shown that in the muscle the myosin-ATPase is most probably also subjected to the inhibitory action of the Mg ions. 6. From data in the literature it is calculated that the liberation of inorganic phosphate during muscular activity takes place at a rate of at least 0.200 mg. P per mg. myosin per minute. 7. From the results of the present study it is found that the myosin in the muscle can liberate inorganic phosphate from ATP at a rate of at most 0.003 mg. P per mg. myosin per minute. 8. It is concluded therefore that myosin-ATPase cannot be responsible for the liberation of the main part of the phosphate in contracting muscle, and therefore cannot have the rôle in muscular metabolism ascribed to it in recent hypotheses and discussions.

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The inhibitory action of relaxing-factor preparations on the myofibrillar adenosine triphosphatase

Biochemical Journal ◽

10.1042/bj0770262 ◽

1960 ◽

Vol 77 (2) ◽

pp. 262-271 ◽

Cited By ~ 32

Author(s):

G. D. Baird ◽

S. V. Perry

Keyword(s):

Inhibitory Action ◽

Adenosine Triphosphatase ◽

Relaxing Factor

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The nature and properties of the inducible sodium-plus-potassium ion-dependent adenosine triphosphatase in the gills of eels (Anguilla anguilla) adapted to fresh water and sea water

Biochemical Journal ◽

10.1042/bj1440069 ◽

1974 ◽

Vol 144 (1) ◽

pp. 69-75 ◽

Cited By ~ 50

Author(s):

John R. Sargent ◽

Alison J. Thomson

Keyword(s):

Fresh Water ◽

Adenosine Triphosphatase ◽

Sea Water ◽

Specific Activity ◽

Sodium Dodecyl ◽

Anguilla Anguilla ◽

Gill Tissue ◽

Sodium Deoxycholate ◽

Enzyme Preparations ◽

Protein Components

1. Gill tissue from eels adapted to fresh water or to sea water was disrupted in 0.32m-sucrose containing 0.1% (w/v) sodium deoxycholate and the subcellular distribution of (Na++K+)-dependent adenosine triphosphatase was determined. 2. About 70% of the recovered enzyme was in a fraction sedimenting between 225000gav.-min and 6000000gav.-min; the specific activities of enzymes from tissues of freshwater and seawater eels were 16 and 51 μmol of phosphate/h per mg of protein respectively. 3. The enzymes from gills of freshwater and seawater eels were indistinguishable on the basis of a number of parameters. These included phosphorylation by [γ-32P]ATP, the binding of [3H]ouabain, the extent to which bound [3H]ouabain was displaced by increasing concentrations of KCl and pH optima. 4. Electrophoresis on polyacrylamide gels in sodium dodecyl sulphate showed that enzyme preparations from both sources had an identical number of protein components. 5. The higher specific activity of (Na++K+)-dependent adenosine triphosphatase from tissue of seawater eels was accompanied by increased amounts of two protein components. One of these proteins retained 32P after treatment of the enzyme with [γ-32P]ATP and had mol.wt. 97000; the other component was a glycoprotein with mol.wt. approx. 46000. 6. The results are discussed in terms of the nature of the transepithelial NaCl pumps in the gills of freshwater and seawater fish.

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Molecular Structure of the Outermost Cell Wall Component of Spirillum Serpens

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100079218 ◽

1977 ◽

Vol 35 ◽

pp. 342-343

Author(s):

Wah Chiu ◽

David Grano

Keyword(s):

Molecular Structure ◽

Cell Wall ◽

Outer Membrane ◽

Gel Electrophoresis ◽

Electron Microscopic Examination ◽

Electron Microscopic ◽

Cell Wall Component ◽

Protein Components ◽

Exposure Technique ◽

Structural Aspects

The periodic structure external to the outer membrane of Spirillum serpens VHA has been isolated by similar procedures to those used by Buckmire and Murray (1). From SDS gel electrophoresis, we have found that the isolated fragments contain several protein components, and that the crystalline structure is composed of a glycoprotein component with a molecular weight of ∽ 140,000 daltons (2). Under an electron microscopic examination, we have visualized the hexagonally-packed glycoprotein subunits, as well as the bilayer profile of the outer membrane. In this paper, we will discuss some structural aspects of the crystalline glycoproteins, based on computer-reconstructed images of the external cell wall fragments.The specimens were prepared for electron microscopy in two ways: negatively stained with 1% PTA, and maintained in a frozen-hydrated state (3). The micrographs were taken with a JEM-100B electron microscope with a field emission gun. The minimum exposure technique was essential for imaging the frozen- hydrated specimens.

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Localization of Adenosine Triphosphatase Activity in the Phleom of Nicotiana Tabacum

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100094401 ◽

1972 ◽

Vol 30 ◽

pp. 230-231

Author(s):

James Cronshaw ◽

Jamison E. Gilder

Keyword(s):

Plasma Membrane ◽

Atpase Activity ◽

Adenosine Triphosphatase ◽

Higher Plants ◽

High Surface ◽

Root Tip ◽

Animal Cells ◽

Physiological Processes ◽

Tip Cells ◽

Atpase Localization

Adenosine triphosphatase (ATPase) activity has been shown to be associated with numerous physiological processes in both plants and animal cells. Biochemical studies have shown that in higher plants ATPase activity is high in cell wall preparations and is associated with the plasma membrane, nuclei, mitochondria, chloroplasts and lysosomes. However, there have been only a few ATPase localization studies of higher plants at the electron microscope level. Poux (1967) demonstrated ATPase activity associated with most cellular organelles in the protoderm cells of Cucumis roots. Hall (1971) has demonstrated ATPase activity in root tip cells of Zea mays. There was high surface activity largely associated with the plasma membrane and plasmodesmata. ATPase activity was also demonstrated in mitochondria, dictyosomes, endoplasmic reticulum and plastids.

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Visualisation of E. coli ribosomal RNA in situ by electron spectroscopic imaging and image averaging

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100122733 ◽

1992 ◽

Vol 50 (1) ◽

pp. 466-467

Author(s):

Daniel Beniac ◽

George Harauz

Keyword(s):

Ribosomal Rna ◽

Three Dimensional ◽

Spectroscopic Imaging ◽

Electron Spectroscopic Imaging ◽

E Coli ◽

Microscopical Technique ◽

Quantitative Image ◽

Dimensional Reconstruction ◽

Image Averaging ◽

Protein Components

The structures of E. coli ribosomes have been extensively probed by electron microscopy of negatively stained and frozen hydrated preparations. Coupled with quantitative image analysis and three dimensional reconstruction, such approaches are worthwhile in defining size, shape, and quaternary organisation. The important question of how the nucleic acid and protein components are arranged with respect to each other remains difficult to answer, however. A microscopical technique that has been proposed to answer this query is electron spectroscopic imaging (ESI), in which scattered electrons with energy losses characteristic of inner shell ionisations are used to form specific elemental maps. Here, we report the use of image sorting and averaging techniques to determine the extent to which a phosphorus map of isolated ribosomal subunits can define the ribosomal RNA (rRNA) distribution within them.

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