Identification of the major phosphorylation sites for protein kinase C in kit/stem cell factor receptor in vitro and in intact cells
Blume-Jensen, Peter ; Wernstedt, Christer ; Heldin, Carl-Henrik and Rönnstrand, Lars LU
(1995)
In Journal of Biological Chemistry
270(23).
p.14192-14200
- Abstract
- The c-kit-encoded tyrosine kinase receptor for stem cell factor (Kit/SCFR) is crucial for the development of hematopoietic cells, melanoblasts, and germ cells. Ligand stimulation of Kit/SCFR leads to receptor dimerization and autophosphorylation on tyrosine residues. We recently showed, that protein kinase C (PKC) acts in an SCF-stimulated negative feedback loop, which controls Kit/SCFR tyrosine kinase activity and modulates the cellular responses to SCF (Blume-Jensen, P., Siegbahn, A., Stabel, S., Heldin, C.-H., and Ronnstrand, L. (1993) EMBO J. 12, 4199-4209). We present here the identification of the major phosphorylation sites for PKC in Kit/SCFR. Two serine residues in the kinase insert, Ser-741 and Ser-746, are PKC-dependent... (More)
- The c-kit-encoded tyrosine kinase receptor for stem cell factor (Kit/SCFR) is crucial for the development of hematopoietic cells, melanoblasts, and germ cells. Ligand stimulation of Kit/SCFR leads to receptor dimerization and autophosphorylation on tyrosine residues. We recently showed, that protein kinase C (PKC) acts in an SCF-stimulated negative feedback loop, which controls Kit/SCFR tyrosine kinase activity and modulates the cellular responses to SCF (Blume-Jensen, P., Siegbahn, A., Stabel, S., Heldin, C.-H., and Ronnstrand, L. (1993) EMBO J. 12, 4199-4209). We present here the identification of the major phosphorylation sites for PKC in Kit/SCFR. Two serine residues in the kinase insert, Ser-741 and Ser-746, are PKC-dependent phosphorylation sites in vivo and account for all phosphorylation by PKC in vitro. Together they comprise more than 60% of the total SCF-stimulated receptor phosphorylation in living cells and 85-90% of its phosphorylation in resting cells. Two additional serine residues, Ser-821 close to the major tyrosine autophosphorylation site in the kinase domain and Ser-959 in the carboxyl terminus are SCF-stimulated PKC-dependent phosphorylation sites. However, they are not phosphorylated directly by PKC-alpha in vitro. Both specific receptor tyrosine autophosphorylation and specific receptor-associated phosphatidylinositide 3'-kinase activity was increased approximately 2-fold in response to SCF in PAE cells stably expressing Kit/SCFR(S741A/S746A). Furthermore, the kinase activity of Kit/SCFR(S741A/S746A) toward an exogenous substrate was increased, which was reflected as a decreased Km and an increased Vmax, in accordance with the negative regulatory role of PKC on Kit/SCFR signaling. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1783980
- author
- Blume-Jensen, Peter
; Wernstedt, Christer
; Heldin, Carl-Henrik
and Rönnstrand, Lars
LU
- publishing date
- 1995
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Base SequenceHematopoietic Cell Growth Factors/pharmacologyMolecular Sequence Data*NaphthalenesPhosphatidylinositol 3-KinasesPhosphorylationPhosphotransferases (Alcohol Group Acceptor)/metabolismPolycyclic Compounds/pharmacologyProtein Kinase C/*physiologyProto-Oncogene Proteins/*metabolismProto-Oncogene Proteins c-kitReceptor Protein-Tyrosine Kinases/*metabolismReceptors, Colony-Stimulating Factor/*metabolismSignal TransductionStem Cell FactorTetradecanoylphorbol Acetate/pharmacologyTransfection
- in
- Journal of Biological Chemistry
- volume
- 270
- issue
- 23
- pages
- 14192 - 14200
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- scopus:0029034468
- ISSN
- 1083-351X
- DOI
- 10.1074/jbc.270.23.14192
- language
- English
- LU publication?
- no
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Experimental Clinical Chemistry (013016010)
- id
- d1c32b53-0d30-4afc-bb11-732e86bfdee9 (old id 1783980)
- alternative location
- http://www.jbc.org/content/270/23/14192.full
- date added to LUP
- 2016-04-04 07:48:08
- date last changed
- 2021-01-03 08:09:01
@article{d1c32b53-0d30-4afc-bb11-732e86bfdee9,
abstract = {{The c-kit-encoded tyrosine kinase receptor for stem cell factor (Kit/SCFR) is crucial for the development of hematopoietic cells, melanoblasts, and germ cells. Ligand stimulation of Kit/SCFR leads to receptor dimerization and autophosphorylation on tyrosine residues. We recently showed, that protein kinase C (PKC) acts in an SCF-stimulated negative feedback loop, which controls Kit/SCFR tyrosine kinase activity and modulates the cellular responses to SCF (Blume-Jensen, P., Siegbahn, A., Stabel, S., Heldin, C.-H., and Ronnstrand, L. (1993) EMBO J. 12, 4199-4209). We present here the identification of the major phosphorylation sites for PKC in Kit/SCFR. Two serine residues in the kinase insert, Ser-741 and Ser-746, are PKC-dependent phosphorylation sites in vivo and account for all phosphorylation by PKC in vitro. Together they comprise more than 60% of the total SCF-stimulated receptor phosphorylation in living cells and 85-90% of its phosphorylation in resting cells. Two additional serine residues, Ser-821 close to the major tyrosine autophosphorylation site in the kinase domain and Ser-959 in the carboxyl terminus are SCF-stimulated PKC-dependent phosphorylation sites. However, they are not phosphorylated directly by PKC-alpha in vitro. Both specific receptor tyrosine autophosphorylation and specific receptor-associated phosphatidylinositide 3'-kinase activity was increased approximately 2-fold in response to SCF in PAE cells stably expressing Kit/SCFR(S741A/S746A). Furthermore, the kinase activity of Kit/SCFR(S741A/S746A) toward an exogenous substrate was increased, which was reflected as a decreased Km and an increased Vmax, in accordance with the negative regulatory role of PKC on Kit/SCFR signaling.}},
author = {{Blume-Jensen, Peter and Wernstedt, Christer and Heldin, Carl-Henrik and Rönnstrand, Lars}},
issn = {{1083-351X}},
keywords = {{Base SequenceHematopoietic Cell Growth Factors/pharmacologyMolecular Sequence Data*NaphthalenesPhosphatidylinositol 3-KinasesPhosphorylationPhosphotransferases (Alcohol Group Acceptor)/metabolismPolycyclic Compounds/pharmacologyProtein Kinase C/*physiologyProto-Oncogene Proteins/*metabolismProto-Oncogene Proteins c-kitReceptor Protein-Tyrosine Kinases/*metabolismReceptors; Colony-Stimulating Factor/*metabolismSignal TransductionStem Cell FactorTetradecanoylphorbol Acetate/pharmacologyTransfection}},
language = {{eng}},
number = {{23}},
pages = {{14192--14200}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
series = {{Journal of Biological Chemistry}},
title = {{Identification of the major phosphorylation sites for protein kinase C in kit/stem cell factor receptor in vitro and in intact cells}},
url = {{http://dx.doi.org/10.1074/jbc.270.23.14192}},
doi = {{10.1074/jbc.270.23.14192}},
volume = {{270}},
year = {{1995}},
}