Calcimedins: purification and characterization from chicken gizzard and rat and bovine livers.
A procedure for the simultaneous extraction and purification of four calcimedins from chicken gizzard, rat liver, and bovine liver is described. These proteins bind to hydrophobic resins in a calcium-dependent manner similar to calmodulin and troponin C. The four calcimedins purified had molecular weights 67,000 (67K), 35,000 (35K), 33,000 (33K), and 30,000 (30K) as determined by SDS polyacrylamide gel electrophoresis. Their ability to bind calcium was demonstrated using the Hummel-Dreyer method. Their tissue concentration ranged between 1-4 mg/100 g wet weight in the three tissues studied. During gel filtration, calcimedins 67K and 35K, had Rf (Ve-Vo/Vt-Vo) values of 0.46 and 0.74, respectively, indicating monomeric structure. However, the 33K and 30K calcimedins had Rf values of 0.26 (molecular weights greater than 90,000) suggesting that they occur as subunit complexes in their native state. Antibodies raised against the 67K and 35K calcimedins showed cross reactivity suggesting possible common origin. However, peptide mapping studies showed that they are independent proteins with considerable peptide homology. Antibodies to 30/33K calcimedins did not cross-react with either 67K or 35K calcimedins. Moreover, their peptide maps were strikingly different from those of 67K and 35K calcimedins indicating that they are unique. At present, the regulatory function of this group of proteins is not clear. Indirect evidences support the possibility that they are involved in membrane associated events, such as endocytosis and secretion.
D000818:Animals; D017302:Annexins; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D002417:Cattle; D002645:Chickens; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology; D005895:Gizzard, Avian / Q000032:analysis*; D008099:Liver / Q000032:analysis*; D008970:Molecular Weight; D051381:Rats
D017302:Annexins; D002135:Calcium-Binding Proteins; D000939:Epitopes; D002118:Calcium