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Fisheries Science

, Volume 85, Issue 1, pp 127–135 | Cite as

Purification and biochemical characterization of a vitellogenin-like protein from sea urchin

  • Osamu NishimiyaEmail author
  • Yoshihiko Teraoka
  • Takahiro Gotoh
  • Tomoharu Yuhi
  • Ichiro Higuchi
  • Kazuhiro Ura
  • Yasuaki Takagi
Original Article Biology

Abstract

The major yolk protein in sea urchins is a transferrin-like protein. In this report, a new component was detected in gonad extracts of Strongylocentrotus intermedius and Mesocentrotus nudus, which cross-reacts with antiserum against egg yolk proteins. We tentatively named them egg yolk-related proteins siYRP and mnYRP. The siYRP was purified from testis of S. intermedius by ammonium sulfate fractionation, anion exchange chromatography, affinity chromatography and gel filtration. Purified siYRP was > 900 kDa in size. The siYRP purified on SDS-PAGE under reducing conditions gave seven bands, corresponding to 93, 213 and > 250 kDa. Purified mnYRP displayed similar structural characteristics as siYRP. Purified siYRP and mnYRP were identified by tandem mass spectrometry and renamed as siVitellogenin (Vtg)-like and mnVtg-like proteins, respectively. Both Vtg-like proteins were confirmed to be lipoglycoproteins by staining with Sudan black and periodic acid-Schiff. A specific antiserum against the siVtg-like protein was raised in rabbit. Antiserum against the siVtg-like protein immunostained siVtg-like and mnVtg-like proteins. Immunochemical methods using the antiserum revealed that siVtg-like and mnVtg-like proteins were present in the ovary, testis and unfertilized eggs of both species. These results indicated that Vtg-like proteins have important physiological functions for gonadal growth and gametogenesis in sea urchins.

Keywords

Strongylocentrotus intermedius Mesocentrotus nudus Egg Yolk protein Large lipid transfer protein superfamily 

Notes

Acknowledgements

We thank Dr. Hiroyuki Munehara from the Usujiri Fisheries Station, Field Science Center for the Northern Biosphere, Hokkaido University, for providing samples and helpful advice. This work was supported by the Science and Technology Research Promotion program for Agriculture, Forestry, Fisheries and Food industry, and The Hokusui Society Foundation. We thank the Edanz Group (http://www.edanzediting.com/ac) for editing a draft of this manuscript.

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Copyright information

© Japanese Society of Fisheries Science 2018

Authors and Affiliations

  1. 1.Laboratory of Comparative Physiology, Division of Marine Life Sciences, Faculty of Fisheries SciencesHokkaido UniversityHakodateJapan
  2. 2.South Ehime Fisheries Research CenterEhime UniversityAinanJapan

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