- 139 views
- 87 downloads
Isolation and characterization of calcium binding glycoproteins of cardiac sarcolemmal vesicles
-
- Author(s) / Creator(s)
-
Two major Ca2(+)-binding glycoproteins Mr 120,000 and 100,000 were isolated from 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid -solubilized bovine heart sarcolemma membrane. Peroxidase-conjugated concanavalin A and wheat germ agglutinin lectins bind strongly to the isolated 120- and 100-kDa glycoproteins. Treatment with endoglycosidase F resulted in conversion of the 120-kDa glycoprotein to a form migrating at about 97 kDa. Treatment of the 100-kDa band with endoglycosidase F produced form of about 80 kDa. Endoglycosidase H digestion removes only 5% of the mass of both glycoproteins. the carbohydrate structure of both glycoproteins, is therefore, predicted to be at least 75% complex structure and 25% high mannose or hybrid structure. The 120- and 100-kDa glycoproteins are the major Ca2(+)-binding proteins in the sarcolemma membranes. Intact and endoglycosidase-treated glycoproteins bind 45Ca2+ as analyzed by a 45Ca2+ overlay technique. Using polyclonal antibodies, the 120- and 100-kDa glycoproteins were identified in muscle plasma membranes (ventricles, atria, and uterus smooth muscle). They were, however, not present in non-muscle tissues such as pancreas, liver, and kidney. The 120- and 100-kDa glycoproteins appear to be homologous molecules as judged by their similar V8 protease peptide maps, cross-reactivity with polyclonal antibody, and other physicochemical properties.
-
- Date created
- 1990-01-01
-
- Subjects / Keywords
-
- Animals
- Cattle
- Acetylglucosaminidase
- Concanavalin A
- Muscle
- Calcium Radioisotopes
- Tissue Distribution
- Molecular Weight
- Chromatography
- Electrophoresis
- Calcium-Binding Proteins
- Glycoside Hydrolases
- Wheat Germ Agglutinins
- Mannosyl-Glycoprotein Endo-Beta-N-Acetylglucosaminidase
- Glycoproteins
- Polyacrylamide Gel
- Serine Endopeptidases
- Peptide Mapping
- Cell Membrane Analysis
-
- Type of Item
- Article (Published)
-
- License
- Attribution 4.0 International