Identification of p90, a Prominent Tyrosine-phosphorylated Protein in Fibroblast Growth Factor-stimulated Cells, as 80K-H

Autor: Y. H. Tan, Chia Bin Siak, Graeme R. Guy, Yoon Pin Lim, Kee Chuan Goh, Xinmin Cao, Siew Hwa Ong
Rok vydání: 1996
Předmět:
Recombinant Fusion Proteins
Blotting
Western
Molecular Sequence Data
Biology
Fibroblast growth factor
Biochemistry
Mice
chemistry.chemical_compound
Animals
Humans
Electrophoresis
Gel
Two-Dimensional
Protein phosphorylation
Amino Acid Sequence
Phosphorylation
Tyrosine
Myristoylated Alanine-Rich C Kinase Substrate
Phosphotyrosine
Molecular Biology
Mammals
Platelet-Derived Growth Factor
Epidermal Growth Factor
Calcium-Binding Proteins
Intracellular Signaling Peptides and Proteins
Membrane Proteins
Tyrosine phosphorylation
3T3 Cells
Cell Biology
Fibroblast growth factor receptor 4
Fibroblasts
Fibroblast growth factor receptor 3
Phosphoproteins
Molecular biology
Fusion protein
chemistry
Fibroblast Growth Factor 1
Electrophoresis
Polyacrylamide Gel
Fibroblast Growth Factor 2
Glucosidases
Subcellular Fractions
Zdroj: Journal of Biological Chemistry. 271:5832-5838
ISSN: 0021-9258
DOI: 10.1074/jbc.271.10.5832
Popis: Tyrosine phosphorylation of cellular proteins occurs rapidly upon treatment of fibroblasts with acidic or basic fibroblast growth factors (aFGF, bFGF), suggesting a role for protein phosphorylation in the FGF signaling pathway. Stimulation of Swiss 3T3 cells and MRC-5 fibroblasts with bFGF results in the tyrosine phosphorylation of several proteins, of which the most prominent has been designated as p90. The phosphorylation of p90 is observed within 30 s of treating the cells with FGF but not with other growth factors. Microsequencing of p90 resolved on two-dimensional polyacrylamide gel electrophoresis indicated an N-terminal amino acid sequence which corresponded to a protein previously named as 80K-H. Polyclonal antibodies raised against the predicted C terminus of 80K-H recognized p90 on all Western blots. p90 was found to bind specifically to GRB-2-glutathione S-transferase fusion protein and to be immunoreactive with 80K-H antibody. In addition, anti-phosphotyrosine antibodies immunoprecipitated 80K-H from cell lysates of FGF-stimulated but not from control fibroblasts. The biological function of 80K-H is yet unknown. However, from this study and a previous observation of the obligatory dependence of p90 phosphorylation on FGF receptor occupation, it appears that 80K-H is involved in FGF signaling.
Databáze: OpenAIRE
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