Localization of ras antigenicity in rat hepatocyte plasma membrane and rough endoplasmic reticulum fractions
- Universite de Montreal, Quebec (Canada)
We have examined the antigenicity of plasma membrane (PM) and rough microsomal (RM) fractions from rat liver using anti-ras monoclonal antibodies 142-24EO5 and Y13-259 and immunochemistry as well as electron microscope immunocytochemistry. Proteins immunoprecipitated with monoclonal antibody 142-24E05 were separated using single-dimensional gradient-gel electrophoresis. The separated proteins were then blotted onto nitrocellulose sheets and incubated with (alpha-32P)GTP. Radioautograms of blots indicated the presence of specific 21.5- and 22-kDa labeled proteins in the PM fraction. A 23.5-kDa (alpha-{sup 32}P) GTP-binding protein was detected in immunoprecipitates of both PM and RM fractions. Monoclonal antibody Y13-259 reacted only with the 21.5-kDa (alpha-{sup 32}P) GTP-binding protein in the plasma membrane fraction. When anti-ras monoclonal antibody 142-24E05 and the immunogold technique were applied to membrane fractions using a preembedding immunocytochemical method, specific labeling was observed in association with both vesicular structures and membrane sheets in the PM fraction but only with electron-dense vesicular structures in the RM fraction. Thus ras antigenicity is associated with hepatocyte plasma membranes and ras-like antigenicity is probably associated with vesicular (secretory/endocytic) elements in both plasma membrane and rough microsomal preparations.
- OSTI ID:
- 6152563
- Journal Information:
- Experimental Cell Research; (USA), Vol. 192:1; ISSN 0014-4827
- Country of Publication:
- United States
- Language:
- English
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PROTEINS
BIOLOGICAL LOCALIZATION
ANTIGEN-ANTIBODY REACTIONS
AUTORADIOGRAPHY
CELL MEMBRANES
ELECTROPHORESIS
ENDOPLASMIC RETICULUM
LIVER CELLS
MICROSOMES
MONOCLONAL ANTIBODIES
PHOSPHORUS 32
ANIMAL CELLS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
ISOTOPES
LIGHT NUCLEI
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ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANOIDS
PHOSPHORUS ISOTOPES
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550201* - Biochemistry- Tracer Techniques