Calcium binding protein in squid brain: biochemical similarity to the 28,000-Mr vitamin D-dependent calcium binding protein (calbindin-D28k).
A calcium binding protein that is biochemically similar to vertebrate 28,000-Mr vitamin D-dependent calcium binding protein (calbindin-D28k) has been purified from squid brain. Squid brain calbindin was found to have an isoelectric point of 5.0, was heat stable up to 60 degrees C, and showed increased electrophoretic mobility in the presence of chelator. Amino acid analysis revealed a high content of glutamic and aspartic acids and a low level of methionine, histidine, and tyrosine, a finding similar but not identical to the composition of vertebrate calbindin-D28k. The molecular weight of the squid protein, determined by Ferguson plot analysis of data obtained from sodium dodecyl sulfate-gel electrophoresis, was calculated to be 25,700, as compared with 27,800 for rat renal calbindin. Immunocytochemical analysis demonstrated immunoreactive protein in a selected population of neurons and fibers in several areas of the molluscan nervous system. This study represents the first purification from an invertebrate of a calcium binding protein that is biochemically similar to vitamin D-dependent calcium binding protein. These results demonstrate that calbindin, although not identical in vertebrates and cephalopods, may be phylogenetically conserved in structure. The restricted distribution of immunoreactive calbindin in both the cephalopod and mammalian brain suggests that the function of neuronal calbindin may also be conserved in evolution.
Christakos
S
S
Department of Biochemistry, University of Medicine and Dentistry of New Jersey, Newark 07103.
Malkowitz
L
L
Sori
A
A
Sperduto
A
A
Feldman
S C
SC
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001923:Brain Chemistry; D002135:Calcium-Binding Proteins / Q000032:analysis*; D002845:Chromatography; D049832:Decapodiformes / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D006651:Histocytochemistry; D006358:Hot Temperature; D007124:Immunoenzyme Techniques; D007526:Isoelectric Point; D008970:Molecular Weight; D009420:Nervous System / Q000032:analysis*; D011489:Protein Denaturation; D064030:S100 Calcium Binding Protein G / Q000032:analysis*
D000596:Amino Acids; D002135:Calcium-Binding Proteins; D064030:S100 Calcium Binding Protein G