Tyrosine kinase inhibition affects type 1 angiotensin ii receptor internalization

B. N. Becker, S. Kondo, Jiankang Chen, R. C. Harris

Research output: Contribution to journalArticle

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Abstract

Growth factor receptors activate tyrosine kinases and undergo endocytosis. Recent data suggest that tyrosine kinase inhibition can affect growth factor receptor internalization. The type 1 angiotensin II receptor (AT1R) which is a G-protein-coupled receptor, also activates tyrosine kinases and undergoes endocytosis. Thus, we examined whether tyrosine kinase inhibition affected AT1R internalization. To verify protein tyrosine phosphorylation, both LLCPKCI4 cells expressing rabbit AT1R (LLCPK(AT)1(R)) and cultured rat mesangial cells (MSC) were treated with angiotensin II (Ang II) [1-100 nM] then solubilized and immunoprecipitated with antiphosphotyrosine antisera. Immunoblots of these samples demonstrated that Ang II stimulated protein tyrosine phosphorylation in both cell types. Losartan [1 μM], an AT1R antagonist, inhibited Ang II-stimulated protein tyrosine phosphorylation. LLCPK(AT1R) cells displayed specific 125I-Ang II binding at apical (AP) and basolateral (BL)membranes, and both AP and BL AT1R activated tyrosine phosphorylation. LLCPK(AT)1(R) cells, incubated with genistein (Gen) [200 μM] or tyrphostin B-48 (TB-48) [50 μM], were assayed for acid-resistant specific 125I-Ang II binding, a measure of Ang II internalization. Both Gen (n=7) and TB-48 (n=3) inhibited AP 125I-Ang II internalization (80 ± 7% inhibition; p < 0.025 vs. control). Neither compound affected BL internalization. TB-1, a non-tyrosine kinase-inhibiting tyrphostin, did not affect AP 125I-Ang II endocytosis (n=3), suggesting that the TB-48 effect was specific for tyrosine kinase inhibition. Incubating MSC with Gen (n=5) or herbimycin A [150 ng/ml] (n=4) also inhibited MSC 125I-Ang II internalization (82 ± 11% inhibition; p<0.005 vs. control). Thus, tyrosine kinase inhibition prevented Ang II internalization in MSC and selectively decreased AP Ang II internalization in LLCPK(AT1R) cells suggesting that AP AT1R in LLCPK(AT1R) cells and MSC AT1R have similar endocytic phenotypes, and tyrosine kinase activity may play a role in AT1R internalization.

Original languageEnglish (US)
Pages (from-to)975-993
Number of pages19
JournalJournal of Receptor and Signal Transduction Research
Volume19
Issue number6
DOIs
StatePublished - Jan 1 1999
Externally publishedYes

Fingerprint

Angiotensin Type 1 Receptor
Angiotensins
Angiotensin II
Protein-Tyrosine Kinases
Mesangial Cells
Tyrphostins
Phosphorylation
Tyrosine
Genistein
Endocytosis
Growth Factor Receptors
Proteins
Losartan
G-Protein-Coupled Receptors
Rats
Immune Sera
Phosphotransferases
Cells
Rabbits

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Tyrosine kinase inhibition affects type 1 angiotensin ii receptor internalization. / Becker, B. N.; Kondo, S.; Chen, Jiankang; Harris, R. C.

In: Journal of Receptor and Signal Transduction Research, Vol. 19, No. 6, 01.01.1999, p. 975-993.

Research output: Contribution to journalArticle

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