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Activation of protein kinase C in permeabilized human neuroblastoma SH-SY5Y cells

Larsson, Christer LU ; Saermark, Torben; Mau, Soren and Simonsson, Per LU (1992) In Journal of Neurochemistry 59(2). p.644-651
Abstract
The activation of protein kinase C was investigated in digitonin-permeabilized human neuroblastoma SH-SY5Y cells by measuring the phosphorylation of the specific protein kinase C substrate myelin basic protein4-14. The phosphorylation was inhibited by the protein kinase C inhibitory peptide PKC19-36 and was associated to a translocation of the enzyme to the membrane fractions of the SH-SY5Y cells. 1,2-Dioctanoyl-sn-glycerol had no effect on protein kinase C activity unless the calcium concentration was raised to concentrations found in stimulated cells (above 100 nM). Calcium in the absence of other activators did not stimulate protein kinase C. Phorbol 12-myristate 13-acetate was not dependent on calcium for the activation or the... (More)
The activation of protein kinase C was investigated in digitonin-permeabilized human neuroblastoma SH-SY5Y cells by measuring the phosphorylation of the specific protein kinase C substrate myelin basic protein4-14. The phosphorylation was inhibited by the protein kinase C inhibitory peptide PKC19-36 and was associated to a translocation of the enzyme to the membrane fractions of the SH-SY5Y cells. 1,2-Dioctanoyl-sn-glycerol had no effect on protein kinase C activity unless the calcium concentration was raised to concentrations found in stimulated cells (above 100 nM). Calcium in the absence of other activators did not stimulate protein kinase C. Phorbol 12-myristate 13-acetate was not dependent on calcium for the activation or the translocation of protein kinase C. The induced activation was sustained for 10 min, and thereafter only a small net phosphorylation of the substrate could be detected. Calcium or dioctanoylglycerol, when applied alone, only caused a minor translocation, whereas in combination a marked translocation was observed. Arachidonic acid (10 microM) enhanced protein kinase C activity in the presence of submaximal concentrations of calcium and dioctanoylglycerol. Quinacrine and p-bromophenacyl bromide did not inhibit calcium- and dioctanoylglycerol-induced protein kinase C activity at concentrations which are considered to be sufficient for phospholipase A2 inhibition. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Protein kinase C, Diacylglycerol, Calcium, Arachidonic acid, SHSY5Y cells
in
Journal of Neurochemistry
volume
59
issue
2
pages
644 - 651
publisher
Wiley-Blackwell
external identifiers
  • pmid:1378489
  • scopus:0026753831
ISSN
1471-4159
DOI
10.1111/j.1471-4159.1992.tb09418.x
language
English
LU publication?
yes
id
97474191-50bf-4b8a-9746-e96599864568 (old id 1106157)
date added to LUP
2008-08-04 09:45:57
date last changed
2017-01-01 06:39:41
@article{97474191-50bf-4b8a-9746-e96599864568,
  abstract     = {The activation of protein kinase C was investigated in digitonin-permeabilized human neuroblastoma SH-SY5Y cells by measuring the phosphorylation of the specific protein kinase C substrate myelin basic protein4-14. The phosphorylation was inhibited by the protein kinase C inhibitory peptide PKC19-36 and was associated to a translocation of the enzyme to the membrane fractions of the SH-SY5Y cells. 1,2-Dioctanoyl-sn-glycerol had no effect on protein kinase C activity unless the calcium concentration was raised to concentrations found in stimulated cells (above 100 nM). Calcium in the absence of other activators did not stimulate protein kinase C. Phorbol 12-myristate 13-acetate was not dependent on calcium for the activation or the translocation of protein kinase C. The induced activation was sustained for 10 min, and thereafter only a small net phosphorylation of the substrate could be detected. Calcium or dioctanoylglycerol, when applied alone, only caused a minor translocation, whereas in combination a marked translocation was observed. Arachidonic acid (10 microM) enhanced protein kinase C activity in the presence of submaximal concentrations of calcium and dioctanoylglycerol. Quinacrine and p-bromophenacyl bromide did not inhibit calcium- and dioctanoylglycerol-induced protein kinase C activity at concentrations which are considered to be sufficient for phospholipase A2 inhibition.},
  author       = {Larsson, Christer and Saermark, Torben and Mau, Soren and Simonsson, Per},
  issn         = {1471-4159},
  keyword      = {Protein kinase C,Diacylglycerol,Calcium,Arachidonic acid,SHSY5Y cells},
  language     = {eng},
  number       = {2},
  pages        = {644--651},
  publisher    = {Wiley-Blackwell},
  series       = {Journal of Neurochemistry},
  title        = {Activation of protein kinase C in permeabilized human neuroblastoma SH-SY5Y cells},
  url          = {http://dx.doi.org/10.1111/j.1471-4159.1992.tb09418.x},
  volume       = {59},
  year         = {1992},
}