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Isolation and characterization of the receptor for vitellogenin from follicles of the rainbow trout, Oncorhynchus mykiss

Abstract

The isolation and characterization of the receptor for vitellogenin from follicle membranes of the rainbow trout, Oncorhynchus mykiss, is described. Follicle membrane proteins subjected to SDS-polyacrylamide gel electrophoresis and subsequently to either protein staining or ligand blotting with radiolabelled vitellogenin (125iodine-vitellogenin) demonstrated that the vitellogenin receptor has an apparent molecular mass of 200 kD (probably comprising of two 100-kD subunits) under non-reducing conditions. The vitellogenin binding sites were identified as specific receptors: binding was saturable and the binding sites were both tissue specific to follicle membranes and exhibited ligand specificity. Scatchard analyses of specific binding data revealed a single class of binding sites with a high affinity for rainbow trout vitellogenin (K d=8.2·10-9 mol·1-1). Both brown trout, Salmo trutta, vitellogenin and carp, Cyprinus carpio, vitellogenin were able to displace the radiolabelled rainbow trout vitellogenin from its receptor, although they were less effective than rainbow trout vitellogenin.

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Abbreviations

B max :

maximum number of binding sites available

BSA:

bovine serum albumin

bt-VTG:

brown trout vitellogenin

c-VTG:

earp vitellogenin

HEPES:

N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid

K d :

dissociatian constant

NCM:

nitrocellulose membranes

PMSF:

phenylmethylsulphonylfluoride

rt-VTG:

rainbow trout vitellogenin

VTG:

vitellogenin

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Tyler, C.R., Lancaster, P. Isolation and characterization of the receptor for vitellogenin from follicles of the rainbow trout, Oncorhynchus mykiss . J Comp Physiol B 163, 225–233 (1993). https://doi.org/10.1007/BF00261669

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Key words

  • Vitellogenin
  • Receptor
  • Follicle
  • Rainbow trout, Oncorhynchus mykiss