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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
13
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pubmed:dateCreated |
1985-8-7
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pubmed:abstractText |
Sequences termed v-abl, which encode the protein-tyrosine kinase activity of Abelson murine leukemia virus, have been expressed in Escherichia coli as a fusion product (ptabl50 kinase). This fusion protein contains 80 amino acids of SV40 small t and the 403 amino acid protein kinase domain of v-abl. We report here the purification and characterization of this kinase. The purified material contains two proteins (Mr = 59,800 and 57,200), both of which possess sequences derived from v-abl. Overall purification was 3,750-fold, with a 31% yield, such that 117 micrograms of kinase could be obtained from 40 g of E. coli within 6-7 days. The specific kinase activity is over 170 mumol of phosphate min-1 mumol-1, comparable to the most active protein-serine kinases. Kinase activity is insensitive to K+, Na+, Ca2+, Ca2+-calmodulin, cAMP, or cAMP-dependent protein kinase inhibitor. The Km for ATP is dependent on the concentration of the second substrate. GTP can also be used as a phosphate donor. The enzyme can phosphorylate peptides consisting of as few as two amino acids and, at a very low rate, free tyrosine. Incubation of the kinase with [gamma-32P]ATP results in incorporation of 1.0 mol of phosphate/mol of protein. This reaction, however, cannot be blocked by prior incubation with unlabeled ATP. Incubation of 32P-labeled kinase with either ADP or ATP results in the synthesis of [32P]ATP. This suggests the phosphotyrosine residue on the Abelson kinase contains a high energy phosphate bond.
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pubmed:grant | |
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 Triphosphate,
http://linkedlifedata.com/resource/pubmed/chemical/Angiotensin II,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Viral,
http://linkedlifedata.com/resource/pubmed/chemical/Edetic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/Magnesium,
http://linkedlifedata.com/resource/pubmed/chemical/Manganese,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Kinases,
http://linkedlifedata.com/resource/pubmed/chemical/Protein-Tyrosine Kinases,
http://linkedlifedata.com/resource/pubmed/chemical/Tyrosine
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
5
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pubmed:volume |
260
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pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
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pubmed:pagination |
8070-7
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:2989275-Abelson murine leukemia virus,
pubmed-meshheading:2989275-Adenosine Triphosphate,
pubmed-meshheading:2989275-Angiotensin II,
pubmed-meshheading:2989275-Cell Transformation, Viral,
pubmed-meshheading:2989275-DNA, Viral,
pubmed-meshheading:2989275-Edetic Acid,
pubmed-meshheading:2989275-Escherichia coli,
pubmed-meshheading:2989275-Kinetics,
pubmed-meshheading:2989275-Leukemia Virus, Murine,
pubmed-meshheading:2989275-Magnesium,
pubmed-meshheading:2989275-Manganese,
pubmed-meshheading:2989275-Molecular Weight,
pubmed-meshheading:2989275-Osmolar Concentration,
pubmed-meshheading:2989275-Phosphorylation,
pubmed-meshheading:2989275-Protein Kinases,
pubmed-meshheading:2989275-Protein-Tyrosine Kinases,
pubmed-meshheading:2989275-Tyrosine
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pubmed:year |
1985
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pubmed:articleTitle |
Purification and characterization of a protein-tyrosine kinase encoded by the Abelson murine leukemia virus.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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