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

Author:
Nishimiya, Osamu, Teraoka, Yoshihiko, Gotoh, Takahiro, Yuhi, Tomoharu, Higuchi, Ichiro, Ura, Kazuhiro, Takagi, Yasuaki
Source:
Fisheries science 2019 v.85 no.1 pp. 127-135
ISSN:
0919-9268
Subject:
Mesocentrotus, Strongylocentrotus, Sudan dyes, affinity chromatography, ammonium sulfate, animal ovaries, anion exchange chromatography, antiserum, eggs, fractionation, gametogenesis, gel chromatography, immunochemistry, polyacrylamide gel electrophoresis, rabbits, staining, tandem mass spectrometry, testes, yolk proteins
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.
Agid:
6275972