Neutralization of MHV-A59 by soluble recombinant receptor glycoproteins.
The interaction of viruses with specific receptors is an important determinant of viral tissue tropism and species specificity. Our goals are to understand how mouse hepatitis virus (MHV) recognizes its cellular receptor, MHVR, and how post-binding interactions with this receptor influence viral fusion and entry. Murine cells express a variety of cell surface molecule in the biliary glycoprotein (Bgp) family that are closely related to the MHVR. When these proteins are expressed at high levels in cell culture, they function as MHV receptors. We used a baculovirus expression system to produce soluble recombinant murine Bgp receptors in which the transmembrane and cytoplasmic domains have been replaced with a six-histidine tag. The soluble glycoproteins were purified to apparent homogeneity and shown to react with antisera to the native receptor. We compared the virus neutralizing activities of various soluble receptor glycoproteins. Soluble MHVR [sMHVR(1-4)] had 10-20 fold more virus neutralizing activity the soluble protein derived from the Bgp1b glycoprotein [sBgp1b(1-4)], from MHV-resistant SJL mice. The sMHVR(1-4) glycoprotein was 60-100 fold more active than a truncated receptor molecule containing only the first two immunoglobulin-like domains, sMHVR(1,2). The observation that sMHVR lacking domains 3 and 4 neutralizes MHV-A59 very poorly suggests that these domains may influence virus binding or subsequent steps associated with neutralization.
Department of Microbiology, University of Colorado, Denver 80262, USA.
B D
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Zelus
D R
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Wessner
G S
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Dveksler
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KV
Holmes
D000818:Animals; D015703:Antigens, CD; D015815:Cell Adhesion Molecules; D006023:Glycoproteins / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D006517:Murine hepatitis virus / Q000378:metabolism*; D011991:Receptors, Virus / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012995:Solubility
D015703:Antigens, CD; C069847:CD66 antigens; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins