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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
5
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pubmed:dateCreated |
1986-7-7
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pubmed:abstractText |
The Malachite Green method for determination of inorganic phosphate (Pi) (Itaya K. & Ui, M. (1966) Clin. Chim. Acta 14, 361-366) was modified to measure Pi in the range of 0.2-15 nmol per ml of ATPase reaction mixture. An ATPase reaction mixture is quenched with an equal volume of 0.6 M PCA; the supernatant after centrifugation is mixed with an equal volume of the Malachite Green/molybdate reagent containing 2 g of sodium molybdate, 0.3 g of Malachite Green and 0.5 g of Triton X-100 or Sterox SE in 1 liter of 0.7 M HCl, and the absorbance at 650 nm is then measured after a 35-40 min incubation at 25 degrees C. Owing to the high sensitivity and simplicity of the modified method, the slow time course of myosin ATP hydrolysis in the presence of Mg2+ and the size of initial phosphate burst can be determined accurately using relatively low concentrations of native myosin and its subfragment-1. The phosphate burst size varied with changes in pH, ionic strength, and temperature. A typical value was 0.8-0.9 mol per site in 0.1 M KCl, 10 mM MgCl2, pH 8.0 at 25 degrees C for fresh enzyme preparations.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Adenosine Triphosphatases,
http://linkedlifedata.com/resource/pubmed/chemical/Adenosine Triphosphate,
http://linkedlifedata.com/resource/pubmed/chemical/Myosin Subfragments,
http://linkedlifedata.com/resource/pubmed/chemical/Myosins,
http://linkedlifedata.com/resource/pubmed/chemical/Peptide Fragments,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphates,
http://linkedlifedata.com/resource/pubmed/chemical/Rosaniline Dyes,
http://linkedlifedata.com/resource/pubmed/chemical/malachite green
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pubmed:status |
MEDLINE
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pubmed:month |
May
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pubmed:issn |
0021-924X
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
99
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1465-72
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:2940237-Adenosine Triphosphatases,
pubmed-meshheading:2940237-Adenosine Triphosphate,
pubmed-meshheading:2940237-Animals,
pubmed-meshheading:2940237-Hydrogen-Ion Concentration,
pubmed-meshheading:2940237-Hydrolysis,
pubmed-meshheading:2940237-Myosin Subfragments,
pubmed-meshheading:2940237-Myosins,
pubmed-meshheading:2940237-Osmolar Concentration,
pubmed-meshheading:2940237-Peptide Fragments,
pubmed-meshheading:2940237-Phosphates,
pubmed-meshheading:2940237-Rabbits,
pubmed-meshheading:2940237-Rosaniline Dyes,
pubmed-meshheading:2940237-Spectrophotometry,
pubmed-meshheading:2940237-Temperature
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pubmed:year |
1986
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pubmed:articleTitle |
The initial phosphate burst in ATP hydrolysis by myosin and subfragment-1 as studied by a modified malachite green method for determination of inorganic phosphate.
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pubmed:publicationType |
Journal Article
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