title: The chicken oocyte receptor for yolk precursors as a model for studying the action of receptor-associated protein and lactoferrin.
abstract: Receptor-associated protein (RAP) was originally described as a 39-kDa intracellular protein copurifying with mammalian low density lipoprotein (LDL) receptor-related protein/alpha 2-macroglobulin receptor (LRP/alpha 2MR). RAP has a high affinity for LRP/alpha 2MR and interferes with the receptor's ability to bind a variety of ligands. The laying hen expresses, in a tissue-specific manner, at least four different proteins which belong to the same family of receptors as LRP/alpha 2MR. Here we show that the chicken also produces RAP, so far thought to be expressed only in mammals. Studies on the interaction of recombinant human RAP with the LDL receptor family in the chicken revealed that RAP binds with high affinity to the abundant oocyte receptor for yolk precursors (OVR) as well as to the somatic cell-specific LRP/alpha 2MR. Significantly, RAP interacts with a lower affinity with the LDL receptor, but does not bind to the oocyte-specific form of LRP. Binding of RAP to OVR inhibits the interaction of the receptor with all known physiological ligands, i.e. the yolk precursors very low density lipoprotein, vitellogenin, and alpha 2-macroglobulin. In COS cells transfected with OVR, RAP is internalized and degraded in a concentration-dependent and saturable manner. Lactoferrin, another protein with a high affinity for mammalian LRP/alpha 2MR, also binds to OVR and abolishes its interaction with yolk precursors. Cross-competition experiments show that RAP and lactoferrin recognize sites different from those involved in yolk precursor binding. The availability of pure OVR and LDLR enable us to determine kinetic parameters for the binding of RAP and lactoferrin to these receptors by surface plasmon resonance. Taken together, our results strongly suggest that chicken OVR, which is easily accessible and highly abundant in growing oocytes, represents a superior system for studying mechanistic and structural aspects of the interaction of ligands and modulating proteins with members of the LDL receptor gene family.
authors.affiliation: Department of Molecular Genetics, University of Vienna, Austria.
authors.forename: T
authors.initials: T
authors.lastname: Hiesberger
authors.forename: M
authors.initials: M
authors.lastname: Hermann
authors.forename: L
authors.initials: L
authors.lastname: Jacobsen
authors.forename: S
authors.initials: S
authors.lastname: Novak
authors.forename: R A
authors.initials: RA
authors.lastname: Hodits
authors.forename: H
authors.initials: H
authors.lastname: Bujo
authors.forename: M
authors.initials: M
authors.lastname: Meilinger
authors.forename: M
authors.initials: M
authors.lastname: Hüttinger
authors.forename: W J
authors.initials: WJ
authors.lastname: Schneider
authors.forename: J
authors.initials: J
authors.lastname: Nimpf
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D015153:Blotting, Western; D002645:Chickens; D004527:Egg Proteins / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism; D005260:Female; D026541:Heymann Nephritis Antigenic Complex; D006801:Humans; D007781:Lactoferrin / Q000378:metabolism*; D008077:Lipoproteins, LDL / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009865:Oocytes / Q000502:physiology*; D011498:Protein Precursors / Q000378:metabolism; D051381:Rats; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D011971:Receptors, Immunologic / Q000378:metabolism*; D011973:Receptors, LDL / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014018:Tissue Distribution; D000511:alpha-Macroglobulins / Q000378:metabolism
chemical_list: D004527:Egg Proteins; D026541:Heymann Nephritis Antigenic Complex; D008077:Lipoproteins, LDL; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008562:Membrane Glycoproteins; D011498:Protein Precursors; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D011973:Receptors, LDL; D011993:Recombinant Fusion Proteins; C036053:VLDL receptor; D000511:alpha-Macroglobulins; C030474:vitellogenin receptor; D007781:Lactoferrin