+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

The catalytic domain limits the translocation of protein kinase C alpha in response to increases in Ca2+ and diacylglycerol



The catalytic domain limits the translocation of protein kinase C alpha in response to increases in Ca2+ and diacylglycerol



Biochemical Journal 370(Pt 3): 901-912



Translocation of protein kinase C (PKC) alpha, beta II, delta and epsilon fused to enhanced green fluorescent protein (EGFP) was studied in living neuroblastoma cells by confocal microscopy. Exposure to carbachol elicited transient translocation of PKC alpha-EGFP and beta II-EGFP in most of the cells, PKC delta-EGFP in a few cells and induced sustained translocation of PKC epsilon-EGFP. To monitor levels of Ca(2+) and diacylglycerol and the translocation of PKC in the same cell, the Ca(2+)-sensitive C2 domain, diacylglycerol-sensitive C1 domains and full-length PKC were fused to red, cyan and yellow fluorescent proteins respectively. PKC alpha was translocated a few seconds after the C2 domain, which represents an increase in Ca(2+). This delay was insensitive to removal of the pseudosubstrate in PKC alpha, but the isolated regulatory domain translocated simultaneously with the C2 domain. Translocation of PKC epsilon coincided with the increase in diacylglycerol. Ionomycin induced translocation of PKC alpha and the C2 domain, whereas 1,2-dioctanoylglycerol caused translocation of the C1 domains and PKC epsilon, but not PKC alpha. Experiments with individual C1 domains showed that treatment with carbachol or phorbol 12,13-dibutyrate elicited translocation of PKC alpha C1a, PKC epsilon C1a and PKC epsilon C1b, whereas PKC alpha C1b was largely insensitive to these agents. In contrast with full-length PKC alpha, the regulatory domain of PKC alpha and pseudosubstrate-devoid PKC alpha responded to the carbachol-stimulated increase in diacylglycerol.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 047589565

Download citation: RISBibTeXText

PMID: 12460119

DOI: 10.1042/BJ20021420


Related references

Direct binding of syndecan-4 cytoplasmic domain to the catalytic domain of protein kinase C alpha (PKC alpha) increases focal adhesion localization of PKC alpha. Journal of Biological Chemistry 278(16): 13795-13802, 2003

Induction of filopodia-like protrusions in N1E-115 neuroblastoma cells by diacylglycerol kinase γ independent of its enzymatic activity: potential novel function of the C-terminal region containing the catalytic domain of diacylglycerol kinase γ. Molecular and Cellular Biochemistry 373(1-2): 85-93, 2013

The C-terminal part of diacylglycerol kinase alpha lacking zinc fingers serves as a catalytic domain. Biochemical Journal 318: 583-590, 1996

The plasma membrane translocation of diacylglycerol kinase delta1 is negatively regulated by conventional protein kinase C-dependent phosphorylation at Ser-22 and Ser-26 within the pleckstrin homology domain. Biochemical Journal 382(Pt 3): 957-966, 2004

Subtype-specific translocation of diacylglycerol kinase alpha and gamma and its correlation with protein kinase C. Journal of Biological Chemistry 275(32): 24760-6, 2000

Translocation of Diacylglycerol Kinase j from Cytosol to Plasma Membrane in Response to Activation of G Protein-coupled Receptors and Protein Kinase C. The Journal of Biological Chemistry 280(11): 70-8, 2005

Translocation of diacylglycerol kinase theta from cytosol to plasma membrane in response to activation of G protein-coupled receptors and protein kinase C. Journal of Biological Chemistry 280(11): 9870-9878, 2005

Ultraviolet light irradiation increases cellular diacylglycerol and induces translocation of diacylglycerol kinase in murine keratinocytes. Journal of Investigative Dermatology 99(2): 221-226, 1992

Conformationally constrained analogues of diacylglycerol (DAG). 27. Modulation of membrane translocation of protein kinase C (PKC) isozymes alpha and delta by diacylglycerol lactones (DAG-lactones) containing rigid-rod acyl groups. Journal of Medicinal Chemistry 50(5): 962-978, 2007

Autophosphorylation Suppresses Whereas Kinase Inhibition Augments the Translocation of Protein Kinase Ca in Response to Diacylglycerol. Journal of Biological Chemistry 279(39 S 24 2004): 576-83, 2004

Autophosphorylation suppresses whereas kinase inhibition augments the translocation of protein kinase Calpha in response to diacylglycerol. Journal of Biological Chemistry 279(39): 40576-40583, 2004

Molecular cloning and characterization of protein kinase D: a target for diacylglycerol and phorbol esters with a distinctive catalytic domain. Proceedings of the National Academy of Sciences of the United States of America 91(18): 8572-8576, 1994

Elevated calcium increases ileal villus cell microvillus membrane diacylglycerol content involvement in protein kinase c pkc translocation. Gastroenterology 96(5 PART 2): A92, 1989

Dynamics of diacylglycerol kinase x translocation in living T-cells. Study of the structural domain requirements for translocation and activity. The Journal of Biological Chemistry 277(33): 300-9, 2002

Dynamics of diacylglycerol kinase zeta translocation in living T-cells. Study of the structural domain requirements for translocation and activity. Journal of Biological Chemistry 277(33): 30300-9, 2002