Isolation of a calreticulin-like calcium binding protein from bovine brain.
Intracellular calcium levels are stringently regulated in all cells. The nature of this regulation is incompletely understood, but recent evidence indicates that the endoplasmic reticulum plays an important role in sequestering intracellular calcium. Using methods for isolating both calsequestrin and calreticulin, we have isolated a 58 kDa, high capacity calcium binding protein that exists in microsomes that shift their density in an oxalate-mediated density shift assay. This protein which we call CBP-58 bears similarities to the endoplasmic reticulum protein, calreticulin, in that it has a pI of 4.7 containing approximately 30% glutamate and aspartate, has a high capacity for calcium, and stains blue with the carbocyanine dye, 'Stains-all'. Peptide, amino acid, nucleotide and immunochemical analyses reveal further similarities between CBP-58 and calreticulin, but also some marked differences. Its tissue distribution suggests it is highly enriched in brain versus other tissues. We believe that CBP-58 is a calreticulin-like protein and that differences in the amino acid composition and sequences may reflect species diversity in calreticulin.
Johnson
R J
RJ
Anatomy and Neurobiology, Department, Boston University School of Medicine, MA 02118.
Liu
N G
NG
Fishman
J B
JB
Dixon
J D
JD
Fine
R E
RE
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D001923:Brain Chemistry; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D037282:Calreticulin; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D002848:Chromatography, DEAE-Cellulose; D015342:DNA Probes; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011817:Rabbits; D012689:Sequence Homology, Nucleic Acid
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D015342:DNA Probes; D002118:Calcium