Isolation and purification of calcium-binding protein from electroplax of Electrophorus electricus.
An acidic calcium-binding phosphoprotein has been isolated from a cholinergic tissue, electroplax from Electrophorus electricus. The purification procedures included (NH4)2SO4 fractionation, boiling treatment, ECTEOLA-cellulose chromatography, and gel filtration on Sephadex G-100. Experiments were performed to compare this protein and a calcium-binding protein isolated from mammalian brain, adrenal medulla, and testis. These experiments showed that the two proteins were identical in terms of molecular weight (14 000), calcium-binding dissociation constant (kd=2.1-10(-5) M), electrophoretic mobility at pH 8.7 in 15% polyacrylamide gels, and phosphorus content (1 mol phosphorus per mol protein). In addition, the two proteins had similar amino acid compositions and peptide maps. Although the electroplax protein was not present in eel skeletal muscle, preliminary experiments indicated that small amounts of the protein were present in other eel tissues, namely brain, liver and spleen. These results suggest an identity between the electroplax and mammalian calcium-binding proteins and extend the findind of comparatively large amounts of the protein from mammalian nervous tissue to a cholinergic nervous tissue, electroplax. The close similarity of the proteins suggests a conservation of structure during evolution which may be required to fulfill a role in neuronal function.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D004557:Electric Organ / Q000378:metabolism*; D004593:Electrophorus; D007700:Kinetics; D008970:Molecular Weight; D009124:Muscle Proteins / Q000032:analysis; D009419:Nerve Tissue Proteins / Q000302:isolation & purification* / Q000378:metabolism; D009928:Organ Specificity; D010446:Peptide Fragments / Q000032:analysis; D011485:Protein Binding; D011955:Receptors, Drug; D013045:Species Specificity
D000596:Amino Acids; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; D011955:Receptors, Drug; D002118:Calcium