Isolation and characterization of calcium binding glycoproteins of cardiac sarcolemmal vesicles

Journal Article · Thu Apr 05 04:00:00 EDT 1990 · Journal of Biological Chemistry; (USA)

OSTI ID:6828551

Michalak, M; Fliegel, L; Wlasichuk, K ^[1]

---

1. Univ. of Alberta, Edmonton (Canada)

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.

• OSTI ID: 6828551
• Journal Information: Journal of Biological Chemistry; (USA), Journal Name: Journal of Biological Chemistry; (USA) Vol. 265:10; ISSN JBCHA; ISSN 0021-9258
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AGGLUTININS
ALKALINE EARTH ISOTOPES
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CALCIUM 45
CALCIUM ISOTOPES
CARDIOVASCULAR SYSTEM
CATTLE
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL COMPOSITION
CHROMATOGRAPHY
CONCANAVALIN
DAYS LIVING RADIOISOTOPES
DISTRIBUTION
DOMESTIC ANIMALS
ELECTROPHORESIS
ENZYMES
EVEN-ODD NUCLEI
FRACTIONATION
GLYCOPROTEINS
GLYCOSYL HYDROLASES
HEART
HYDROLASES
INTERMEDIATE MASS NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
MAMMALS
MEMBRANES
MOLECULAR WEIGHT
MUSCLES
MYOCARDIUM
NUCLEI
ORGANIC COMPOUNDS
ORGANS
PEPTIDE HYDROLASES
PROTEINS
RADIOISOTOPES
RUMINANTS
SEPARATION PROCESSES
SERINE PROTEINASES
TISSUE DISTRIBUTION
TRACER TECHNIQUES
VERTEBRATES