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Phosphorylation site-specific inhibition of platelet-derived growth factor beta-receptor autophosphorylation by the receptor blocking tyrphostin AG1296

Kovalenko, Marina ; Rönnstrand, Lars LU ; Heldin, Carl-Henrik ; Loubtchenkov, M ; Gazit, Aviv ; Levitzki, Alexander and Böhmer, Frank D. (1997) In Biochemistry 36(21). p.6260-6269
Abstract
The mechanism of action of AG1296, a potent and specific inhibitor of the platelet-derived growth factor (PDGF) receptor tyrosine kinase [Kovalenko, M., Gazit, A., Bohmer, A., Rorsman, Ch., Ronnstrand, L., Heldin, C.-H., Waltenberger, J., Bohmer, F. D., & Levitzki, A. (1994) Cancer Res. 54, 6106-6114] was investigated. This quinoxalin-type tyrphostin neither interferes with PDGF-BB binding to the PDGF beta-receptor nor has any effect on receptor dimerization. Kinetic analysis of the inhibition was carried out using a synthetic peptide substrate (KY751) corresponding to the sequence around tyrosine 751 autophosphorylation site of the PDGF receptor. It revealed purely competitive inhibition vis-a-vis ATP, mixed competitive inhibition... (More)
The mechanism of action of AG1296, a potent and specific inhibitor of the platelet-derived growth factor (PDGF) receptor tyrosine kinase [Kovalenko, M., Gazit, A., Bohmer, A., Rorsman, Ch., Ronnstrand, L., Heldin, C.-H., Waltenberger, J., Bohmer, F. D., & Levitzki, A. (1994) Cancer Res. 54, 6106-6114] was investigated. This quinoxalin-type tyrphostin neither interferes with PDGF-BB binding to the PDGF beta-receptor nor has any effect on receptor dimerization. Kinetic analysis of the inhibition was carried out using a synthetic peptide substrate (KY751) corresponding to the sequence around tyrosine 751 autophosphorylation site of the PDGF receptor. It revealed purely competitive inhibition vis-a-vis ATP, mixed competitive inhibition vis-a-vis the peptide substrate for the non-activated receptor, and mixed competitive inhibition vis-a-vis both substrates for the activated receptor. Thus, the type of inhibition apparently changes upon receptor activation, indicating conformational changes at the ATP-binding site. The high degree of selectivity for the tyrphostin AG1296 might result from the complex type of interaction with the active center of the receptor as revealed by the kinetic analysis. Dose-response curves for inhibition of the phosphorylation of individual autophosphorylation sites of the PDGF beta-receptor by AG1296 were different, phosphorylation of tyrosine 857 being the most susceptible to inhibition. Thus, phosphorylation of tyrosine 857 in the PDGF receptor kinase domain seems dispensable for partial kinase activation. The findings are discussed in relation to current models of receptor tyrosine kinase activation. (Less)
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published
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keywords
Platelet-Derived Growth Factor/*antagonists &inhibitors/metabolismSwine*Tyrphostins, Adenosine Triphosphate/metabolismAmino Acid SequenceAnimalsBinding, CompetitiveCell LineDimerizationDogsEnzyme Inhibitors/*pharmacologyHumansKineticsMolecular Sequence DataNitriles/*pharmacologyPhosphatidylinositol 3-KinasesPhosphorylationPhosphotransferases (Alcohol Group Acceptor)/metabolismProtein Binding/drug effectsQuinoxalines/*pharmacologyReceptor Protein-Tyrosine Kinases/*antagonists & inhibitors/metabolismReceptors
in
Biochemistry
volume
36
issue
21
pages
6260 - 6269
publisher
The American Chemical Society (ACS)
external identifiers
  • scopus:0342546626
ISSN
0006-2960
DOI
10.1021/bi962553l
language
English
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no
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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)
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944af2e9-1fdc-47f4-af0a-6bb40bc286f4 (old id 1783921)
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http://pubs.acs.org/doi/abs/10.1021/bi962553l
date added to LUP
2016-04-04 07:07:53
date last changed
2020-05-26 15:25:28
@article{944af2e9-1fdc-47f4-af0a-6bb40bc286f4,
  abstract     = {The mechanism of action of AG1296, a potent and specific inhibitor of the platelet-derived growth factor (PDGF) receptor tyrosine kinase [Kovalenko, M., Gazit, A., Bohmer, A., Rorsman, Ch., Ronnstrand, L., Heldin, C.-H., Waltenberger, J., Bohmer, F. D., & Levitzki, A. (1994) Cancer Res. 54, 6106-6114] was investigated. This quinoxalin-type tyrphostin neither interferes with PDGF-BB binding to the PDGF beta-receptor nor has any effect on receptor dimerization. Kinetic analysis of the inhibition was carried out using a synthetic peptide substrate (KY751) corresponding to the sequence around tyrosine 751 autophosphorylation site of the PDGF receptor. It revealed purely competitive inhibition vis-a-vis ATP, mixed competitive inhibition vis-a-vis the peptide substrate for the non-activated receptor, and mixed competitive inhibition vis-a-vis both substrates for the activated receptor. Thus, the type of inhibition apparently changes upon receptor activation, indicating conformational changes at the ATP-binding site. The high degree of selectivity for the tyrphostin AG1296 might result from the complex type of interaction with the active center of the receptor as revealed by the kinetic analysis. Dose-response curves for inhibition of the phosphorylation of individual autophosphorylation sites of the PDGF beta-receptor by AG1296 were different, phosphorylation of tyrosine 857 being the most susceptible to inhibition. Thus, phosphorylation of tyrosine 857 in the PDGF receptor kinase domain seems dispensable for partial kinase activation. The findings are discussed in relation to current models of receptor tyrosine kinase activation.},
  author       = {Kovalenko, Marina and Rönnstrand, Lars and Heldin, Carl-Henrik and Loubtchenkov, M and Gazit, Aviv and Levitzki, Alexander and Böhmer, Frank D.},
  issn         = {0006-2960},
  language     = {eng},
  number       = {21},
  pages        = {6260--6269},
  publisher    = {The American Chemical Society (ACS)},
  series       = {Biochemistry},
  title        = {Phosphorylation site-specific inhibition of platelet-derived growth factor beta-receptor autophosphorylation by the receptor blocking tyrphostin AG1296},
  url          = {http://dx.doi.org/10.1021/bi962553l},
  doi          = {10.1021/bi962553l},
  volume       = {36},
  year         = {1997},
}