Selective protein transport: characterization and solubilization of the phosvitin receptor from chicken oocytes.
Phosvitin (PV), a subunit of a female-specific protein, vitellogenin, binds to oocyte membranes with a KD of 10(-6) M. Binding reaches equilibrium within 30 min after incubation at 25 degrees C. Bound 125I-PV dissociates from the membrane with a t1/2 of 13 h when incubated in buffer. However, when 125I-PV-labeled membranes are incubated in buffer containing 10(-5) M unlabeled PV, 50% of the initially bound 125I-PV dissociates from the membrane within 10 min. These results support the conclusion that PV binds to a membrane-associated receptor. Solubilization studies show that Triton X-100 solubilizes up to 45% of the total membrane-bound 125I-PV. Gel-exclusion chromatography of the solubilized material yields a 500,000 dalton 125 I-PV-containing complex separated from free 125I-PV. The 500,000 dalton complex completely dissociates to yield free 125I-PV when incubated with excess unlabeled PV. However, when incubated with 1) no addition, 2) IgG, or 3) serum albumin, the extent of dissociation is significantly reduced and is consistent with that which would be predicted on the basis of the observed dissociation rate in the absence of unlabeled PV. These results suggest that bound 125I-PV can only be displaced by unlabeled PV. These results also indicate that the 500,000 dalton species is a solubilized PV-receptor complex and that it is possible to solubilize the PV-receptor in an active form.
D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002642:Chick Embryo; D002850:Chromatography, Gel; D004527:Egg Proteins / Q000378:metabolism*; D005260:Female; D007700:Kinetics; D017830:Octoxynol; D009865:Oocytes / Q000378:metabolism*; D010063:Ovum / Q000378:metabolism*; D010774:Phosvitin / Q000378:metabolism*; D011092:Polyethylene Glycols; D011955:Receptors, Drug / Q000302:isolation & purification*; D012995:Solubility; D014819:Vitellogenins / Q000378:metabolism
D004527:Egg Proteins; D011955:Receptors, Drug; D014819:Vitellogenins; D011092:Polyethylene Glycols; D017830:Octoxynol; D010774:Phosvitin