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Journal of Virology, October 2002, p. 10383-10392, Vol. 76, No. 20
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.20.10383-10392.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

A Structural Model of Pestivirus E^rns Based on Disulfide Bond Connectivity and Homology Modeling Reveals an Extremely Rare Vicinal Disulfide

J. P. M. Langedijk,^1,2* P. A. van Veelen,³ W. M. M. Schaaper,¹ A. H. de Ru,³ R. H. Meloen,¹ and M. M. Hulst²

Pepscan Systems Inc., 8203 AB Lelystad,¹ Department of Mammalian Virology, Institute for Animal Science and Health (ID-Lelystad), 8200 AB Lelystad,² Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, NL-2300 RC Leiden, The Netherlands³

Received 30 April 2002/ Accepted 2 July 2002

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.

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* Corresponding author: Mailing address: Pepscan Systems Inc., Edelhertweg 15, P.O. Box 2098, 8203 AB Lelystad, The Netherlands. Phone: 31-320-237209. Fax: 31-320-238120. E-mail: j.p.m.langedijk{at}id.wag-ur.nl.

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Journal of Virology, October 2002, p. 10383-10392, Vol. 76, No. 20
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.20.10383-10392.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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