Purification and partial characterization of vitellogenin from shorthead redhorse (Moxostoma macrolepidotum) and copper redhorse (Moxostoma hubbsi) and detection in plasma and mucus with a heterologous antibody
Three charge isomeric forms of vitellogenin are reported in the blood of estrogen-treated murrel, Channa punctatus on the basis of the results of native and SDS-PAGE, isoelectric focusing, and Ferguson plot analysis. Vitellogenin was induced in adult murrels by exogenous administration of estradiol-17beta and labelled with 32P in vivo. Labelled vitellogenin isolated from other plasma proteins on Ultrogel AcA 34 columns resolve into three bands on native-PAGE. Ferguson plot analysis reveals free electrophoretic mobilities of the three bands as 6.0, 4.5, and 3.7, which are very similar to the isoelectric point values of 5.9, 4.6, and 3.8, respectively. The apparent molecular weight of native murrel vitellogenin is 530 kDa. It consists of a single peptide with a molecular weight of 175 kDa. All the three bands on native PAGE resolves into a single peptide on SDS-PAGE. N-terminal amino acid sequence for murrel vitellogenin peptide is MKAVVLALLL. The protein phosphorus:lipid phosphorus ratio for murrel vitellogenin is 0.9. The total lipid content is 32.8%, consisting of 45% neutral lipids and 30% phospholipids. Phosphatidyl choline is the major phospholipid whereas triglycerides are the major neutral lipids. Results of these analytical analyses indicate that murrel native vitellogenin circulates as three charge isomers; poor in phosphorus but rich in lipids content.