title: Receptor-associated protein in an oviparous species is correlated with the expression of a receptor variant.
abstract: The biosynthesis of proteins containing cysteine-rich domains requires chaperones for their correct folding. For instance, the 39-kDa receptor-associated protein (RAP) aides in the cell-surface targeting of newly synthesized members of the mammalian low density lipoprotein receptor (LDLR) gene family, which contains tandemly arranged clusters of hexacysteine repeats. In the chicken, an LDLR relative with eight such repeats is expressed as two different splice variant forms in cell type-specific fashion (Bujo, H., Lindstedt, K. A., Hermann, M., Mola Dalmau, L., Nimpf, J., and Schneider, W. J. (1995) J. Biol. Chem. 270, 23546-23551). To learn more about evolutionary aspects of RAP, its role in escorting of these different receptor splice variants, and other potential functions, we have extended our studies on the avian LDLR family to RAP. cDNA cloning, determination of tissue expression at both the transcript and the protein level, stable expression in COS cells, and binding studies with chicken RAP revealed that mammalian RAPs have retained many features of the non-amniotic proteins. However, structural details, e.g. the well defined internal triplicate repeats in the chicken protein, have been somewhat diluted during evolution. Interestingly, chicken RAP was found to correlate positively with the expression levels in somatic cells of the larger splice variant of the eight-cysteine repeat receptor, but not with those of the smaller variant, expressed only in germ cells. This is compatible with the possibility that RAP may play a role in receptor biology that could be complementing its function in assisting folding. Chicken RAP in crude extracts of the stable expressor COS cells is able to bind to LDLR relatives in ligand blots without requirement for prior purification of the ligand. Thus, in conjunction with the avian model of massive lipid transport to germ cells, these cells provide a novel comparative system amenable to investigation of the biological functions of RAP.
authors.affiliation: Department of Molecular Genetics, University and Biocenter Vienna, Dr. Bohr Gasse 9/2, A-1030 Vienna, Austria. wjs@mol.univie.ac.at
authors.forename: K A
authors.initials: KA
authors.lastname: Lindstedt
authors.forename: M G
authors.initials: MG
authors.lastname: Mahon
authors.forename: R
authors.initials: R
authors.lastname: Foisner
authors.forename: M
authors.initials: M
authors.lastname: Hermann
authors.forename: J
authors.initials: J
authors.lastname: Nimpf
authors.forename: W J
authors.initials: WJ
authors.lastname: Schneider
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D019556:COS Cells; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002645:Chickens; D018076:DNA, Complementary / Q000235:genetics; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017403:In Situ Hybridization; D026563:LDL-Receptor Related Protein-Associated Protein; D018832:Molecular Chaperones / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011973:Receptors, LDL / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014018:Tissue Distribution
chemical_list: D002352:Carrier Proteins; D018076:DNA, Complementary; D006023:Glycoproteins; D026563:LDL-Receptor Related Protein-Associated Protein; D018832:Molecular Chaperones; D011973:Receptors, LDL