Purification and characterization of a novel incomplete-type vitellogenin protein (VgC) in Sakhalin taimen (Hucho perryi).
A novel, incomplete-type vitellogenin (VgC) and its derived yolk lipovitellin (LvC) were immunologically detected in female serum and egg extracts, respectively, of Sakhalin taimen (Hucho perryi) using a subtype-specific antiserum against LvC of grey mullet (Mugil cephalus). The taimen VgC was purified from the sera of vitellogenic females by a combination of gel filtration, anion exchange, and immunoadsorbent column chromatography. Gel filtration of the purified VgC revealed that it had an apparent native mass of approximately 380 kDa, while the mass of the VgC polypeptide that appeared following SDS-PAGE was estimated to be approximately 140 kDa. An antiserum was raised against the purified VgC and utilized for the development of a subtype-specific immunoassay for VgC. Levels of VgC in the serum of female taimen increased from 25 microg/mL to approximately 1mg/mL, with an increase of GSI. Levels of complete-type Vg and estradiol-17beta (E2) in the serum of E2-administered juvenile taimen increased and reached peak levels similar to those found in vitellogenic females. Although VgC could be induced in the serum of E2-administered taimen, it stayed at levels (35.5-73 microg/mL) lower than those obtained in females. This is the first report on the presence of serum VgC and yolk LvC in a salmonid species; these findings indicate that for Sakhalin taimen, like other highly-evolved teleost species, this minor subtype of Vg is significant in the formation of egg yolk.
Amano
Haruna
H
Faculty of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido 041-8611, Japan.
Mochizuki
Machiko
M
Fujita
Toshiaki
T
Hiramatsu
Naoshi
N
Todo
Takashi
T
Hara
Akihiko
A
D000818:Animals; D004527:Egg Proteins / Q000097:blood / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification*; D004530:Egg Yolk / Q000737:chemistry / Q000276:immunology; D005260:Female; D007118:Immunoassay; D012487:Salmonidae; D013045:Species Specificity; D014819:Vitellogenins / Q000097:blood / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification*
D004527:Egg Proteins; D014819:Vitellogenins; C001549:lipovitellin