Linked Life Data

10867018

Source:http://linkedlifedata.com/resource/pubmed/id/10867018

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
26
pubmed:dateCreated
2000-8-10
pubmed:abstractText
Activation of the serine/threonine kinase, protein kinase D (PKD/PKC mu) via a phorbol ester/PKC-dependent pathway involves phosphorylation events. The present study identifies five in vivo phosphorylation sites by mass spectrometry, and the role of four of them was investigated by site-directed mutagenesis. Four sites are autophosphorylation sites, the first of which (Ser(916)) is located in the C terminus; its phosphorylation modifies the conformation of the kinase and influences duration of kinase activation but is not required for phorbol ester-mediated activation of PKD. The second autophosphorylation site (Ser(203)) lies in that region of the regulatory domain, which in PKC mu interacts with 14-3-3tau. The last two autophosphorylation sites (Ser(744) and Ser(748)) are located in the activation loop but are only phosphorylated in the isolated PKD-catalytic domain and not in the full-length PKD; they may affect enzyme catalysis but are not involved in the activation of wild-type PKD by phorbol ester. We also present evidence for proteolytic activation of PKD. The fifth site (Ser(255)) is transphosphorylated downstream of a PKC-dependent pathway after in vivo stimulation with phorbol ester. In vivo phorbol ester stimulation of an S255E mutant no longer requires PKC-mediated events. In conclusion, our results show that PKD is a multisite phosphorylated enzyme and suggest that its phosphorylation may be an intricate process that regulates its biological functions in very distinct ways.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
30
pubmed:volume
275
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
19567-76
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed-meshheading:10867018-Alanine, pubmed-meshheading:10867018-Alkaline Phosphatase, pubmed-meshheading:10867018-Binding Sites, pubmed-meshheading:10867018-Cell Line, pubmed-meshheading:10867018-Down-Regulation, pubmed-meshheading:10867018-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:10867018-Enzyme Inhibitors, pubmed-meshheading:10867018-Glutamic Acid, pubmed-meshheading:10867018-Humans, pubmed-meshheading:10867018-Indoles, pubmed-meshheading:10867018-Kinetics, pubmed-meshheading:10867018-Maleimides, pubmed-meshheading:10867018-Mass Spectrometry, pubmed-meshheading:10867018-Models, Biological, pubmed-meshheading:10867018-Mutagenesis, Site-Directed, pubmed-meshheading:10867018-Phosphorylation, pubmed-meshheading:10867018-Precipitin Tests, pubmed-meshheading:10867018-Protein Kinase C, pubmed-meshheading:10867018-Recombinant Proteins, pubmed-meshheading:10867018-Signal Transduction, pubmed-meshheading:10867018-Time Factors, pubmed-meshheading:10867018-Transfection, pubmed-meshheading:10867018-Trypsin
pubmed:year
2000
pubmed:articleTitle
Regulation of protein kinase D by multisite phosphorylation. Identification of phosphorylation sites by mass spectrometry and characterization by site-directed mutagenesis.
pubmed:affiliation
Afdeling Biochemie, Faculteit Geneeskunde, Campus Gasthuisberg, Katholieke Universiteit Leuven, Herestraat 49, B-3000 Leuven, Belgium.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't