title: A structural model of pestivirus E(rns) based on disulfide bond connectivity and homology modeling reveals an extremely rare vicinal disulfide.
abstract: E(rns) is a pestivirus envelope glycoprotein and is the only known viral surface protein with RNase activity. E(rns) is a disulfide-linked homodimer of 100 kDa; it is found on the surface of pestivirus-infected cells and is secreted into the medium. In this study, the disulfide arrangement of the nine cysteines present in the mature dimer was established by analysis of the proteolytically cleaved protein. Fragments were obtained after digestion with multiple proteolytic enzymes and subsequently analyzed by liquid chromatography-electrospray ionization mass spectrometry. The analysis demonstrates which cysteine is involved in dimerization and reveals an extremely rare vicinal disulfide bridge of unknown function. With the assistance of the disulfide arrangement, a three-dimensional model was built by homology modeling based on the alignment with members of the Rh/T2/S RNase family. Compared to these other RNase family members, E(rns) shows an N-terminal truncation, a large insertion of a cystine-rich region, and a C-terminal extension responsible for membrane translocation. The homology to mammalian RNase 6 supports a possible role of E(rns) in B-cell depletion.
authors.affiliation: Pepscan Systems Inc., Edelhertweg 15, PO Box 2098, 8203 AB Lelystad, The Netherlands. j.p.m.langedijk@id.wag-ur.nl
authors.forename: J P M
authors.initials: JP
authors.lastname: Langedijk
authors.forename: P A
authors.initials: PA
authors.lastname: van Veelen
authors.forename: W M M
authors.initials: WM
authors.lastname: Schaaper
authors.forename: A H
authors.initials: AH
authors.lastname: de Ru
authors.forename: R H
authors.initials: RH
authors.lastname: Meloen
authors.forename: M M
authors.initials: MM
authors.lastname: Hulst
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D006692:Classical Swine Fever Virus; D004220:Disulfides; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D004220:Disulfides; C466501:E(rns) protein, classic swine fever virus; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D014357:Trypsin