2024-03-29T06:20:18Zhttps://eprints.lib.hokudai.ac.jp/dspace-oai/requestoai:eprints.lib.hokudai.ac.jp:2115/303492022-11-17T02:08:08Zhdl_2115_20049hdl_2115_141Purification of multiple vitellogenins in grey mullet (Mugil cephalus)Amano, HarunaFujita, ToshiakiHiramatsu, NaoshiSawaguchi, SayumiMatsubara, TakahiroSullivan, Craig VHara, Akihiko487Three female specific serum proteins were detected immunologically in the sera of grey mullet (Mugil cephalus) which were named vitellogenin A (VgA), VgB, and VgC, based upon their distinct antigenicity against specific antisera raised against three types of mullet lipovitellins (Lvs). These Vgs were subsequently purified from the serum of estradiol-treated mullet by combining several types of chromatography columns (anion exchanger, hydroxylapatite, immunoadsorbent column, and gel filtration). Purified native VgA, VgB, and VgC exhibited molecular masses of 570, 580, and 335 kDa, respectively. Following, SDS-PAGE, the estimated mass of polypeptide bands evident for VgA and VgB were ~179 kDa and ~175 kDa, respectively; VgC appeared to be ~132 kDa. The two larger Vgs (VgA and VgB) appeared to be phosphorylated, suggesting that these Vgs contain a highly phosphorylated, serine-rich phosvitin (Pv) domain. Furthermore, two discrete Vg-type specific antisera, anti-VgA and anti-VgB, were developed and each generated two precipitin lines against ovary extracts in immunoelectrophoresis, indicating that these Vgs contain additional antigenic yolk protein domains: Lv and β'-component. The small Vg (VgC) appeared to lack a Pv domain because of its low serine content (5.35%) and failure to show positive results in phospho-staining experiments. In conjunction with N-terminal amino acid sequencing analyses of the purified Vgs, our present results have conclusively identified the purified Vg products in grey mullet as typical A-type (VgA), B-type (VgB), and C-type (VgC) Vgs.SpringerJournal Articleapplication/pdfhttp://hdl.handle.net/2115/30349https://eprints.lib.hokudai.ac.jp/dspace/bitstream/2115/30349/1/amano_hara.pdf0025-31621432-1793Marine Biology1526121512252007-11enginfo:doi/10.1007/s00227-007-0768-zThe original publication is available at www.springerlink.comauthor