Tyrosine-phosphorylated caveolin-1: Immunolocalization and molecular characterization

Ryuji Nomura, Toyoshi Fujimoto

研究成果: Article

96 引用 (Scopus)

抄録

Caveolin-1 was discovered as a major substrate for v-Src, but the effect of its tyrosine phosphorylation has not been known. We generated a specific antibody (PY14) to caveolin-1 phosphorylated at tyrosine 14 and studied the significance of the modification. By Western blotting of lysates of v-Src- expressing cells, PY14 recognized not only a 22-kDa band (the position of nonphosphorylated caveolin-1) but bands at 23-24 and 25 kDa. Bands of slower mobility were diminished by dephosphorylation and were also observed for mutant caveolin-1 lacking tyrosine 14. By immunofluorescence microscopy, PY14 did not label normal cells but detected large dots in v-Src-expressing cells. Immunoelectron microscopy revealed that the dots corresponded to aggregated caveolae and/or vesicles of various sizes; besides, the label was observed in intramembrane particle-free areas in the plasma membrane, which appeared to have been formed by fusion of flattened caveolae. A positive reaction with PY14 was found in normal cells after vanadate or pervanadate treatment; it occurred mainly at 22 kDa by Western blotting and was not seen as large dots by immunofluorescence microscopy. Detergent solubility, oligomerization, and association with caveolin-2 were observed similarly for caveolin-1 in normal and v-Src-expressing cells. The results indicate that phosphorylation of caveolin-1 in v-Src-expressing cells occurs at multiple residues and induces flattening, aggregation, and fusion of caveolae and/or caveolae-derived vesicles.

元の言語English
ページ(範囲)975-986
ページ数12
ジャーナルMolecular Biology of the Cell
10
発行部数4
DOI
出版物ステータスPublished - 01-01-1999

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Caveolin 1
Tyrosine
Caveolae
Fluorescence Microscopy
Caveolin 2
Western Blotting
Phosphorylation
Vanadates
Immunoelectron Microscopy
Detergents
Solubility
Cell Membrane
Antibodies

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

これを引用

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Tyrosine-phosphorylated caveolin-1 : Immunolocalization and molecular characterization. / Nomura, Ryuji; Fujimoto, Toyoshi.

:: Molecular Biology of the Cell, 巻 10, 番号 4, 01.01.1999, p. 975-986.

研究成果: Article

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