title: Identification of receptor-interacting regions of vitellogenin within evolutionarily conserved β-sheet structures by using a peptide array.
abstract: Vitellogenesis, a key process in oviparous animals, is characterized by enhanced synthesis of the lipoprotein vitellogenin, which serves as the major yolk-protein precursor. In most oviparous animals, and specifically in crustaceans, vitellogenin is mainly synthesized in the hepatopancreas, secreted to the hemolymph, and taken up into the ovary by receptor-mediated endocytosis. In the present study, localization of the vitellogenin receptor and its interaction with vitellogenin were investigated in the freshwater prawn Macrobrachium rosenbergii. The receptor was immuno-histochemically localized to the cell periphery and around yolk vesicles. A receptor blot assay revealed that the vitellogenin receptor interacts with most known vitellogenin subunits, the most prominent being the 79 kDa subunit. The receptor was, moreover, able to interact with trypsin-digested vitellogenin peptides. By combining a novel peptide-array approach with tandem mass spectrometry, eleven vitellogenin-derived peptides that interacted with the receptor were identified. A 3D model of vitellogenin indicated that four of the identified peptides are N-terminally localized. One of the peptides is homologous to the receptor-recognized site of vertebrate vitellogenin, and assumes a conserved β-sheet structure. These findings suggest that this specific β-sheet region in the vitellogenin N-terminal lipoprotein domain is the receptor-interacting site, with the rest of the protein serving to enhance affinity for the receptor. The conservation of the receptor recognition site in invertebrate and vertebrate vitellogenin might have vast implications for oviparous species reproduction, development, immunity, and pest management.
authors.affiliation: Avram and Stella Goldstein-Goren Department of Biotechnology, Engineering, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, Israel.
authors.forename: Ziv
authors.initials: Z
authors.lastname: Roth
authors.forename: Simy
authors.initials: S
authors.lastname: Weil
authors.forename: Eliahu D
authors.initials: ED
authors.lastname: Aflalo
authors.forename: Rivka
authors.initials: R
authors.lastname: Manor
authors.forename: Amir
authors.initials: A
authors.lastname: Sagi
authors.forename: Isam
authors.initials: I
authors.lastname: Khalaila
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004527:Egg Proteins / Q000737:chemistry / Q000378:metabolism; D019143:Evolution, Molecular; D008024:Ligands; D008969:Molecular Sequence Data; D033541:Palaemonidae / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D040081:Protein Array Analysis; D017433:Protein Structure, Secondary; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism; D014819:Vitellogenins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D004527:Egg Proteins; D008024:Ligands; D010455:Peptides; D011956:Receptors, Cell Surface; D014819:Vitellogenins; C030474:vitellogenin receptor