Characterization of a vitellogenin receptor in white perch (Morone americana).
Receptors for white perch vitellogenin (wVTG) were characterized using wVTG, labeled in vivo with [3H]leucine or in vitro with 125I, and semipurified ovarian membranes. Specific binding of wVTG to the membranes was temperature-dependent, proportional to the amount of membrane, and saturable. Scatchard analyses revealed a single class of binding sites of low maximum binding capacity (MBC; approximately 35 pmol VTG/mg membrane protein) and high affinity (Kd approximately 400 nM), consistent with wVTG levels (540-2700 nM) circulating in maturing females. Ligand blotting revealed a receptor protein of M(r) approximately 157000 and a smaller protein, possibly its degradation product. Striped bass vitellogenin, chicken egg yolk very low density lipoprotein, and suramin displaced wVTG from its receptor, but BSA did not. No change in Kd was noted over the course of vitellogenesis in maturing perch, and MBC increased only slightly very late in the gametogenic cycle. The wVTG bound specifically to membranes prepared from liver, muscle, and mesenteric fat, but not to erythrocyte membranes. The Kd for ovary (394 nM) and liver (345 nM) were similar, but the Kd for muscle (1440 nM) was much higher.
Tao
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Department of Zoology, North Carolina State University, Raleigh, North Carolina 27695, USA.
Berlinsky
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Sullivan
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D000818:Animals; D001667:Binding, Competitive; D015153:Blotting, Western; D004527:Egg Proteins; D004530:Egg Yolk / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D007457:Iodine Radioisotopes; D007700:Kinetics; D008079:Lipoproteins, VLDL / Q000737:chemistry / Q000302:isolation & purification; D008565:Membrane Proteins / Q000378:metabolism; D008970:Molecular Weight; D010053:Ovary / Q000378:metabolism*; D010471:Perches / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
D004527:Egg Proteins; D007457:Iodine Radioisotopes; D008079:Lipoproteins, VLDL; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; C030474:vitellogenin receptor