Identification of a calsequestrin-like protein from sea urchin eggs.
Following studies on calcium transport by isolated smooth endoplasmic reticulum from unfertilized sea urchin eggs (Oberdorf, J. A., Head, J. F., and Kaminer, B. (1986) J. Cell Biol. 102, 2205-2210) we have purified and partially characterized a calsequestrin-like protein from this organelle isolated from eggs from Strongylocentrotus droebachiensis and Arbacia punctulata. Muscle calsequestrin from sarcoplasmic reticulum is well characterized as a calcium storage protein. The egg protein resembles calsequestrin in its behavior in purification steps, electrophoretic mobility, blue staining with Stains-all on polyacrylamide gels, and its calcium binding and amino acid composition. Purification was attained with DEAE-cellulose and hydroxyapatite chromatography. The egg protein Mr of 58,000 in the Laemmli gel system is reduced to 54,000 under Weber-Osborn (neutral) conditions, thus showing a pH dependence in its mobility, although less than occurs with muscle calsequestrins. 25% of its amino acids are acidic and 10% basic. It binds 309 nmol of Ca2+/mg of protein, within the range reported for cardiac calsequestrin. Antigenically, the sea urchin egg protein is related to cardiac calsequestrin capable of binding anti-cardiac calsequestrin antibody.
Oberdorf
J A
JA
Department of Physiology, Boston University School of Medicine, Massachusetts 02118.
Lebeche
D
D
Head
J F
JF
Kaminer
B
B
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002155:Calsequestrin / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D002845:Chromatography; D002848:Chromatography, DEAE-Cellulose; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006882:Hydroxyapatites; D008861:Microsomes / Q000378:metabolism; D009124:Muscle Proteins / Q000302:isolation & purification*; D010063:Ovum / Q000378:metabolism*; D012617:Sea Urchins / Q000378:metabolism
D000596:Amino Acids; D002155:Calsequestrin; D006882:Hydroxyapatites; D009124:Muscle Proteins; D017886:Durapatite; D002118:Calcium