Vitellogenin-derived yolk proteins in a hybrid sturgeon, bester (Huso huso x Acipencer ruthenus): identification, characterization and course of proteolysis during embryogenesis.
Vitellogenin (Vg) and its corresponding yolk protein (YP) products, YP1, YP2 and YP3, were isolated from serum of estrogen-treated hybrid sturgeon (bester; Huso huso X Acipencer ruthenus) and eggs from untreated fish, respectively. Vitellogenin had an apparent molecular mass of 580 kDa and appeared as two major bands corresponding to 180 kDa and 120 kDa after SDS-PAGE. Apparent molecular weights of YP1, YP2 and YP3 were 370 kDa, 88 kDa and 19 kDa, respectively. After SDS-PAGE, YP1 appeared as a main band of 110 kDa, while YP2 was resolved as a single band of 94 kDa and 29 kDa band under non-reducing and reducing conditions, respectively. Yolk protein 3 appeared as a diffuse band corresponding to 16 kDa and two faint bands below 14.4 kDa after SDS-PAGE. However, the 16 kDa band alone was observed after dephosphorylation with alkaline phosphatase. The course of cleavage of yolk proteins in bester embryos and alevins was observed by SDS-PAGE and Western blotting from fertilization onward. After hatching, the main 110 kDa band of YP1 was degraded into smaller peptides during development, while YP2 hardly showed any such structural changes. The amino acid compositions of purified yolk proteins indicated that YP1, YP2 and YP3 were bester lipovitellin, beta-component, and phosvitin, respectively.
Hiramatsu
Naoshi
N
Department of Zoology, College of Agriculture and Life Sciences, North Carolina State University, Raleigh, NC 27695-7617, USA.
Hiramatsu
Kaori
K
Hirano
Kaori
K
Hara
Akihiko
A
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D015153:Blotting, Western; D002850:Chromatography, Gel; D004527:Egg Proteins / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D005399:Fishes / Q000196:embryology* / Q000378:metabolism; D006824:Hybridization, Genetic; D008970:Molecular Weight; D011233:Precipitin Tests; D011485:Protein Binding; D014819:Vitellogenins / Q000737:chemistry*
D000596:Amino Acids; D004527:Egg Proteins; D014819:Vitellogenins