Purification and identification of a second form of vitellogenin from ascites of medaka (Oryzias latipes) treated with estrogen

Estrogen treatment of medaka leads to accumulation of ascites, in which vitellogenin (Vg) and choriogenins (precursors to vitelline envelope) are abundant. Besides those female‐specific proteins, we detected a new component in ascites that cross‐reacts with antiserum against egg yolk proteins. We te...

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Published in:The Journal of experimental zoology 2002-12, Vol.293 (7), p.726-735
Main Authors: Shimizu, Munetaka, Fujiwara, Yumi, Fukada, Haruhisa, Hara, Akihiko
Format: Article
Language:English
Subjects:
Amino Acids - analysis
Animals
Antibody Specificity
Ascitic Fluid - chemistry
Blotting, Western
choriogenins
Egg Proteins - chemistry
Egg Proteins - immunology
egg yolk-related protein
Electrophoresis, Polyacrylamide Gel
Estrogens - administration & dosage
Estrogens - pharmacology
Female
Freshwater
Immunodiffusion
Male
Molecular Weight
Oryzias - physiology
Oryzias latipes
Vitellogenins - analogs & derivatives
Vitellogenins - chemistry
Vitellogenins - immunology
Vitellogenins - isolation & purification
YRP protein
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Summary:Estrogen treatment of medaka leads to accumulation of ascites, in which vitellogenin (Vg) and choriogenins (precursors to vitelline envelope) are abundant. Besides those female‐specific proteins, we detected a new component in ascites that cross‐reacts with antiserum against egg yolk proteins. We tentatively named it egg yolk‐related protein (YRP). YRP was purified from ascites by hydroxylapatite chromatography followed by gel filtration. Purified YRP had a molecular mass of 460kDa in intact state while 570kDa for Vg. The molecular weight of purified YRP on SDS‐PAGE under both reducing and nonreducing conditions was 130kDa. YRP was confirmed to be a lipoglycophosphoprotein by staining with Sudan black, periodic acid‐Schiff (PAS) and methyl green. Amino acid composition of YRP resembled that of Vg except for a relatively low content of serine. A specific antiserum against YRP was raised in a rabbit. Antiserum against YRP specifically immunostained its antigen but not Vg or choriogenins. YRP was detected as a female‐specific protein in serum of breeding medaka. The antiserum also cross‐reacted with a band at 29kDa in egg extracts, which is not immunoreactive to antiserum against Vg. These data show that YRP is a precursor to some egg yolk proteins with differing antigenicity from Vg (Hamazaki et al. '87). We thus conclude that YRP is a second form of medaka Vg and rename YRP as Vg 2 while formerly reported Vg as Vg 1. J. Exp. Zool. 293:726–735, 2002. © 2002 Wiley‐Liss, Inc.
ISSN:0022-104X
1097-010X
DOI:10.1002/jez.10178