RPE cell surface proteins in normal and dystrophic rats
Membrane-bound proteins in plasma membrane enriched fractions from cultured rat RPE were analyzed by two-dimensional gel electrophoresis. Membrane proteins were characterized on three increasingly specific levels. Total protein was visualized by silver staining. A maximum of 102 separate proteins were counted in silver-stained gels. Glycoproteins were labeled with 3H-glucosamine or 3H-fucose and detected by autoradiography. Thirty-eight fucose-labeled and 61-71 glucosamine-labeled proteins were identified. All of the fucose-labeled proteins were labeled with glucosamine-derived radioactivity. Proteins exposed at the cell surface were labeled by lactoperoxidase-catalyzed radioiodination prior to preparation of membranes for two-dimensional analysis. Forty separate 125I-labeled surface proteins were resolved by two-dimensional electrophoresis/autoradiography. Comparison with the glycoprotein map showed that a number of these surface labeled proteins were glycoproteins. Two-dimensional maps of total protein, fucose-labeled, and glucosamine-labeled glycoproteins, and 125I-labeled surface proteins of membranes from dystrophic (RCS rdy-p+) and normal (Long Evans or RCS rdy+p+) RPE were compared. No differences in the total protein or surface-labeled proteins were observed. However, the results suggest that a 183K glycoprotein is more heavily glycosylated with glucosamine and fucose in normal RPE membranes as compared to membranes from dystrophic RPE.
- OSTI ID:
- 5844651
- Journal Information:
- Invest. Ophthalmol. Visual Sci.; (United States), Journal Name: Invest. Ophthalmol. Visual Sci.; (United States) Vol. 2; ISSN IOVSD
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
AMINES
ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
CARBOHYDRATES
CELL CONSTITUENTS
CELL CULTURES
CELL MEMBRANES
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
GLUCOPROTEINS
GLUCOSAMINE
HEXOSAMINES
HEXOSES
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IODINE ISOTOPES
ISOTOPES
MAMMALS
MEMBRANES
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NUCLEI
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ORGANIC COMPOUNDS
PROTEINS
RADIOISOTOPES
RATS
RODENTS
SACCHARIDES
VERTEBRATES