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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
27
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pubmed:dateCreated |
1993-10-20
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pubmed:abstractText |
Signal transduction for tumor necrosis factor-alpha and interleukin-1 involves sphingomyelin hydrolysis to ceramide and stimulation of a ceramide-activated serine/threonine protein kinase (Mathias, S., Younes, A., Kan, C., Orlow, I., Joseph, C., and Kolesnick, R. (1993) Science 259, 519-522). Kinase activity is detected by phosphorylation of a 19-amino acid peptide derived from the sequence surrounding Thr669 of the epidermal growth factor receptor. Thr669 is contained within a -Pro-Leu-Thr-Pro- motif, which conforms to a known recognition sequence for the proline-directed class of serine/threonine protein kinases. The present studies used peptides with single-site amino acid substitutions within this sequence to assess substrate recognition by ceramide-activated protein kinase. Substitution of alanine for the C-terminal but not the N-terminal proline reduced kinase activity by 80%. Similarly, substitution of basic residues for the leucine residue reduced kinase activity by 90%. Substitution of acidic residues for leucine, or its removal, also markedly reduced kinase activity. Surprisingly, addition of a leucine residue between threonine and the C-terminal proline enhanced kinase activity 3-4 fold. The Vmax(app) of the enzyme toward the control peptide containing -Pro-Leu-Thr-Pro- (200 +/- 11 pmol of peptide phosphorylated/min/mg of membrane protein) was enhanced 2.3-fold by ceramide. However, ceramide had no effect on the Km (2.0 +/- 0.4 mM). Membranes containing ceramide-activated protein kinase showed minimal activity toward peptides derived from substrates for casein kinase II, S6 kinase, protein kinase C, and cAMP-dependent protein kinase, but possessed substantial activity toward a calmodulin kinase substrate. However, activities toward these substrates were not enhanced by ceramide. These results suggest that ceramide-activated protein kinase may be a member of the proline-directed class of protein kinases and display specificity for -Leu-Thr-Pro- as a minimal substrate recognition motif.
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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/Alanine,
http://linkedlifedata.com/resource/pubmed/chemical/Ceramides,
http://linkedlifedata.com/resource/pubmed/chemical/Oligopeptides,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphopeptides,
http://linkedlifedata.com/resource/pubmed/chemical/Proline,
http://linkedlifedata.com/resource/pubmed/chemical/Protein-Serine-Threonine Kinases
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pubmed:status |
MEDLINE
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pubmed:month |
Sep
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
25
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pubmed:volume |
268
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
20002-6
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pubmed:dateRevised |
2007-11-15
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pubmed:meshHeading |
pubmed-meshheading:8376361-Alanine,
pubmed-meshheading:8376361-Amino Acid Sequence,
pubmed-meshheading:8376361-Ceramides,
pubmed-meshheading:8376361-Chromatography, High Pressure Liquid,
pubmed-meshheading:8376361-Humans,
pubmed-meshheading:8376361-Intracellular Membranes,
pubmed-meshheading:8376361-Kinetics,
pubmed-meshheading:8376361-Leukemia, Promyelocytic, Acute,
pubmed-meshheading:8376361-Microsomes,
pubmed-meshheading:8376361-Oligopeptides,
pubmed-meshheading:8376361-Phosphopeptides,
pubmed-meshheading:8376361-Proline,
pubmed-meshheading:8376361-Protein-Serine-Threonine Kinases,
pubmed-meshheading:8376361-Substrate Specificity,
pubmed-meshheading:8376361-Tumor Cells, Cultured
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pubmed:year |
1993
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pubmed:articleTitle |
Substrate recognition by ceramide-activated protein kinase. Evidence that kinase activity is proline-directed.
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pubmed:affiliation |
Laboratory of Signal Transduction, Memorial Sloan-Kettering Cancer Center, New York, New York 10021.
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pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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