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Identification of four ovarian receptor proteins that bind vitellogenin but not other homologous plasma lipoproteins in the rainbow trout, Oncorhynchus mykiss

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Abstract

Membrane proteins from ovarian follicles, testis and somatic tissues of rainbow trout, Oncorhynchus mykiss, were extracted by ultracentrifugation, separated on sodium dodecyl sulphate gels and isolated on polyvinyl difluoride membranes. Vitellogenin receptor proteins were visualised using protein staining and hybridisation with 125I-vitellogenin Four follicle-membrane proteins, with molecular masses of 220, 210, 110 and 100 kDa, showed a strong affinity for vitellogenin and were specific to the ovary. Other homologous lipoproteins (very low density lipoprotein, low density lipoprotein and high density lipoprotein) had a very limited ability to displace 125I-vitellogenin from its receptor, indicating that the ovarian receptor proteins were fairly specific for vitellogenin. Proteins with an affinity for very low density lipoprotein and low density lipoprotein were visualised in liver, spleen and muscle, eluting on sodium dodecyl sulphate gels with molecular masses of about 150 kDa. Peptides generated from trypsin digests of the receptor proteins with a high affinity for vitellogenin showed sequence homology with receptors in the lipoprotein family, including a sequence that is believed to act as the internalisation signal [Phe-Asp-Asn-Phe-Tyr-] and, a sequence identity with the recently characterised chicken vitellogenin/very low density lipoprotein receptor [Ser-Glu-Leu-Tyr-Glu-Pro-Ala-]. Together, the ligand blotting and peptide sequence data support the contention that the four ovarian membrane proteins isolated are receptor proteins specific for vitellogenin and they do not bind other plasma lipoproteins to any significant degree.

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Abbreviations

BSA :

bovine serum albumin

HDL :

high density lipoprotein

LDL :

low-density lipoprotein

HPLC :

high performance liquid chromatograph

PVDF :

polyvinylidene difluoride

RIA :

radioimmunoassay

rt-VTG :

rainbow trout vitellogenin

SDS :

sodium dodecyl sulphate

VLDL :

very low density lipoprotein

VTG :

vitellogenin

VRP-1,-2,-3 or -4 :

vitellogenin receptor proteins

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Communicated by H. Huddart

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Tyler, C.R., Lubberink, K. Identification of four ovarian receptor proteins that bind vitellogenin but not other homologous plasma lipoproteins in the rainbow trout, Oncorhynchus mykiss . J Comp Physiol B 166, 11–20 (1996). https://doi.org/10.1007/BF00264634

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