Identification of phosphoproteins associated with maintenance of transformed state in temperature-sensitive Rous sarcoma-virus infected cells by proteomic analysis

Journal Article · Fri Jul 07 04:00:00 EDT 2006 · Biochemical and Biophysical Research Communications

DOI:https://doi.org/10.1016/j.bbrc.2006.04.183· OSTI ID:20854346

Yamaoka, Kazuko ^[1]; Imajoh-Ohmi, Shinobu ^[2]; Fukuda, Hiroyuki ^[2]; Akita, Yoshiko ^[3]; Kurosawa, Keiko ^[2]; Yamamoto, Yukio ^[4]; Sanai, Yutaka ^[1]

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1. Laboratory of Glycobiology, Tokyo Metropolitan Institute of Medical Science, Tokyo (Japan)
2. Institute of Medical Science, University of Tokyo, Tokyo (Japan)
3. Laboratory of Mouse Model for Human Heritable Disease, Tokyo Metropolitan Institute of Medical Science, Tokyo (Japan)
4. Tumor Therapy Project, Tokyo Metropolitan Institute of Medical Science, Tokyo (Japan)

To identify phosphotyrosine-containing proteins essential for maintaining the transformed state, we studied the tyrosine phosphorylation profile of temperature-sensitive mutant of Rous sarcoma virus, tsNY68, infected cells (68N7). Shifting the temperature from 39 {sup o}C (nonpermissive) to 32 {sup o}C (permissive) markedly increased the expression of phosphotyrosine-containing cell membrane proteins of {approx}40 kDa, as assessed by SDS-PAGE. Membrane and nuclear proteins were separated by two-dimensional gel electrophoresis and immunoblotted with anti-phosphotyrosine antibody. Proteins showing temperature-dependent changes in phosphorylation profile were subjected to in-gel digestion with trypsin and analyzed by mass spectrometry. Five proteins were identified: heterogeneous nuclear ribonucleoprotein (hnRNP) A3, hnRNP A2, annexin II, phosphoglycerate mutase 1, and triosephosphate isomerase 1. hnRNP A3 was phosphorylated at serine residues and had both serine and tyrosine phosphorylated sites. These results suggest an important complementary role for proteomics in identifying molecular abnormalities associated with tumor progression that may be attractive candidates for tumor diagnosis.

• OSTI ID: 20854346
• Journal Information: Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 3 Vol. 345; ISSN 0006-291X; ISSN BBRCA9
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
ANTIBODIES
CELL MEMBRANES
DIAGNOSIS
DIGESTION
ELECTROPHORESIS
GELS
MASS SPECTROSCOPY
MUTANTS
NEOPLASMS
ONCOGENIC VIRUSES
PHOSPHOPROTEINS
PHOSPHORYLATION
SERINE
TEMPERATURE DEPENDENCE
TRYPSIN
TYROSINE