Isolation and properties of a sarcoplasmic calcium-binding protein from crayfish.
The sarcoplasmic calcium-binding protein from crayfish muscle has been purified to homogeneity. The protein has a molecular weight of 44000, as determined by sedimentation equilibrium and Sephadex chromatography. It dissociates in the presence of sodium dodecyl sulfate, 8 M urea, or, after succinylation, into two subunits of 22000 molecular weight. The protein is free of carbohydrate and phosphorus but contains 4 g-atoms of calcium/44000 at a free calcium concentration of 0.1 muM. Approximately 45% of the polypeptide backbone appears to be alpha-helical. The amino acid composition reveals a high proportion of alanine and acidic amino acids, a normal content of aromatic amino acids, and the absence of histidine. The isoelectric point, as determined by isoelectric focusing, is 5.1. The protein contains a free threonyl NH2 terminal. Two thiols react rapidly in the native protein, six in the calcium-free form. Immunochemically, there is no difference between the protein from tail, claw, and heart muscle. In these three crayfish tissues, the concentrations of calcium-binding protein, as determined by rocket immunoelectrophoresis, are markedly different: 2.73 g/kg in tail, 0.72 in claw, and 0.073 in heart muscle. A functional analogy with the parvalbumins of vertebrates can be postulated.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D003400:Astacoidea / Q000378:metabolism*; D001665:Binding Sites; D002118:Calcium / Q000032:analysis / Q000378:metabolism*; D002942:Circular Dichroism; D005779:Immunodiffusion; D046911:Macromolecular Substances; D008970:Molecular Weight; D009124:Muscle Proteins / Q000302:isolation & purification / Q000378:metabolism; D009132:Muscles / Q000032:analysis; D009206:Myocardium / Q000032:analysis; D009928:Organ Specificity; D011485:Protein Binding; D011487:Protein Conformation; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D046911:Macromolecular Substances; D009124:Muscle Proteins; D002118:Calcium