RRC ID 41517
Author Kashiwagi K, Shirai Y, Kuriyama M, Sakai N, Saito N.
Title Importance of C1B domain for lipid messenger-induced targeting of protein kinase C.
Journal J Biol Chem
Abstract The molecular mechanisms by which arachidonic acid (AA) and ceramide elicit translocation of protein kinase C (PKC) were investigated. Ceramide translocated epsilonPKC from the cytoplasm to the Golgi complex, but with a mechanism distinct from that utilized by AA. Using fluorescence recovery after photobleaching, we showed that, upon treatment with AA, epsilonPKC was tightly associated with the Golgi complex; ceramide elicited an accumulation of epsilonPKC which was exchangeable with the cytoplasm. Stimulation with ceramide after AA converted the AA-induced Golgi complex staining to one elicited by ceramide alone; AA had no effect on the ceramide-stimulated localization. Using point mutants and deletions of epsilonPKC, we determined that the epsilonC1B domain was responsible for the ceramide- and AA-induced translocation. Switch chimeras, containing the C1B from epsilonPKC in the context of deltaPKC (delta(epsilonC1B)) and vice versa (epsilon(deltaC1B)), were generated and tested for their translocation in response to ceramide and AA. delta(epsilonC1B) translocated upon treatment with both ceramide and AA; epsilon(deltaC1B) responded only to ceramide. Thus, through the C1B domain, AA and ceramide induce different patterns of epsilonPKC translocation and the C1B domain defines the subtype specific sensitivity of PKCs to lipid second messengers.
Volume 277(20)
Pages 18037-45
Published 2002-5-17
DOI 10.1074/jbc.M111761200
PII S0021-9258(20)85303-4
PMID 11877428
MeSH Animals Arachidonic Acid / metabolism CHO Cells COS Cells Ceramides / metabolism Cricetinae Cytoplasm / drug effects Cytoplasm / metabolism Golgi Apparatus / drug effects Golgi Apparatus / metabolism Green Fluorescent Proteins HeLa Cells Humans Isoenzymes / chemistry Isoenzymes / genetics Isoenzymes / metabolism* Luminescent Proteins / genetics Point Mutation Protein Kinase C / chemistry Protein Kinase C / genetics Protein Kinase C / metabolism* Protein Kinase C-delta Protein Kinase C-epsilon Spectrometry, Fluorescence Structure-Activity Relationship Tetradecanoylphorbol Acetate / pharmacology
IF 4.238
Times Cited 66
Human and Animal Cells COS-7(RCB0539) CHO-K1(RC0285)