Purification and characterization of a calsequestrin-like calcium-binding protein from carp (Cyprinus carpio) sarcoplasmic reticulum.
1. A calsequestrin-like calcium-binding protein was purified from carp sarcoplasmic reticulum by column chromatographies using DEAE-cellulose and Butyl-Toyopearl 650S. 2. The mol. wt was estimated to be 50 kDa, which was larger than that of rabbit calsequestrin (42 kDa). 3. Carp calsequestrin-like protein bound Ca2+ with a higher affinity (apparent Kd = 400 microM) and lower capacity (25 mol/mol) compared with rabbit calsequestrin (1 mM and 40-50 mol/mol, respectively). 4. Anti-carp calsequestrin-like protein rabbit antiserum reacted with rabbit calsequestrin in immunoblotting analysis. 5. Carp calsequestrin-like protein was rich in acidic amino acids, as was rabbit calsequestrin.
Watabe
S
S
Laboratory of Marine Biochemistry, Faculty of Agriculture, University of Tokyo, Japan.
Ushio
H
H
Hashimoto
K
K
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002155:Calsequestrin / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002347:Carps / Q000378:metabolism*; D007120:Immunochemistry; D008970:Molecular Weight; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D013045:Species Specificity
D000596:Amino Acids; D002135:Calcium-Binding Proteins; D002155:Calsequestrin