Biochemical and Immunochemical Characterization of Two Discrete Vitellogenin Proteins and Their Derived Lipovitellins in the Inshore Hagfish (Eptatretus burgeri)

Abstract

Vitellogenesis has been extensively studied in oviparous vertebrates, including teleost fishes, while not much is known with regard to jawless hagfishes, modern representatives of the most primitive vertebrate class. This study aimed to characterize vitellogenin (Vtg) and yolk protein (YP) in the inshore hagfish (Eptatretus burgeri) as an initial step to understand vitellogenesis in this species. A putative Vtg fraction was purified from the serum of female hagfish by combinations of hydroxylapatite and ion-exchange chromatography, followed by gel filtration. The purified fraction appeared to contain two distinct Vtgs (Vtg1 and Vtg2) and exhibited biochemical properties resembling those previously reported for teleost Vtgs; these appeared to be female-specific serum proteins and high-molecular-weight proteins in gel filtration (~505 kDa as the mixture fraction of both Vtgs) and in SDS-PAGE (Vtg1 and Vtg2; ~210 kDa and ~195 kDa, respectively). A major YP was also purified from hagfish eggs by combinations of hydroxylapatite chromatography and gel filtration; the apparent native mass of the purified YP was unusually large (> 669 kDa). The purified YP consisted of four polypeptides in SDS-PAGE; the peptide pattern indicated that it consisted of two lipovitellins (Lv1 and Lv2) giving rise to two sets of heavy chains (~116 kDa and ~106 kDa, respectively) and two light chains (~32 kDa and ~28 kDa, respectively). Additional immunological analysis, Nterminal amino acid sequencing and cDNA cloning firmly confirmed the precursor-product relationship between hagfish Vtgs and Lvs.

Resolving the complexity of vitellogenins and their receptors in the tetraploid Atlantic salmon (Salmo salar): Ancient origin of the phosvitin‐less VtgC in chondrichthyean fishes

“…Phylogenetic analyses of vertebrate Vtgs suggested that the phosvitinless VtgC is a neo-functional product of the second whole-genome duplication that occurred subsequent to the divergence of jawless (Agnathans) and jawed (Gnathostomes) vertebrates (Finn & Kristoffersen, 2007;Prowse & Byrne, 2012). The presence of phosvitin in the Vtgs of invertebrates (such as the mosquito), agnathan fishes, and the Indonesian coelacanth (Latimeria menadoensis) (Canapa et al, 2012;Chen, Sappington, & Raikhel, 1997;Nishimiya et al, 2014;Sharrock et al, 1992)-indicates that this domain was lost after the divergence of ray-finned (Actinopterygians) and lobe-finned (Sarcopterygians) fishes.…”