Analysis of ras oncogene products by two-dimensional gel electrophoresis: evidence for protein families with distinctive molecular forms

• Published in: Biochimica et biophysica acta Vol. 866; no. 4; pp. 204 - 215
• Main Authors: FUHRER, J. P, DEBIASI, F, COOPER, H. L, SCHLOM, J
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier 05.05.1986; North-Holland
• Subjects: Amino Acid Sequence; Amino Acids - analysis; Animals; Biological and medical sciences; Cell Transformation, Neoplastic - metabolism; Chemical Precipitation; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; Genes. Genome; Humans; Mice; Molecular and cellular biology; Molecular genetics; Neoplasm Proteins - analysis; Neoplasm Proteins - immunology; Neoplasms - analysis; Oncogene Protein p21(ras); Radioimmunoassay; Transfection; Vertebrata; Molecular form; Cell line; Mammalia; Gene product; C-Onc gene; Multigene family; Two dimensional electrophoresis
• ISSN: 0006-3002; 1878-2434

Protein products of the ras family of oncogenes were immunoprecipitated by an anti-p21 monoclonal antibody, separated by two-dimensional gel electrophoresis and subsequently detected by western immunoblot analysis using the same anti-p21 monoclonal antibody as a probe. Using this method, a 21 kDa oncogene protein (p21) was detected and characterized in cell lines containing Harvey (Ha), Kirsten (Ki), neuroblastoma (N), or cellular (proto) ras genes. The ras gene products from all cell types occurred with multiple forms differing in size, charge or in both parameters. Transforming ras oncogene proteins occurred in easily identifiable groups that were different from each other in molecular weight and charge, were distinctive for each ras gene type and were different from cellular ras equivalents. Similar, but not identical, family groups occurred in different cell types containing the same oncogene. The reproducible occurrence of unpredicted p21 forms suggests that previously unreported post-translational processing steps may be associated with the synthesis of certain oncogene products. This immunoprecipitation/two-dimensional gel/western blot technique is a simple method for the identification and characterization of p21 gene products.