HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Supplementary table Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Sissoëff, L. Articles by Tuffereau, C. Search for Related Content PubMed PubMed Citation Articles by Sissoëff, L. Articles by Tuffereau, C. Agricola Articles by Sissoëff, L. Articles by Tuffereau, C.

Stable trimerization of recombinant rabies virus glycoprotein ectodomain is required for interaction with the p75^NTR receptor

Ludmilla Sissoëff^1,

Mohamed Mousli^1,,

¹ Laboratoire de Virologie Moléculaire et Structurale, UMR 2472 CNRS-INRA, 91198 Gif-sur-Yvette, France
² Plateforme de Biophysique des Macromolécules et de leurs Interactions, Institut Pasteur, 75015 Paris, France

Correspondence
Christine Tuffereau
christine.tuffereau{at}vms.cnrs-gif.fr

Native rabies virus glycoprotein (RVGvir) is a trimeric, membrane-anchored protein that has been shown to interact with the p75^NTR neurotrophin receptor. In order to determine if the RVG trimeric oligomerization state is required for its binding with p75^NTR, different soluble recombinant molecules containing the entire RVG ectodomain (RVGect) were expressed alone or fused at its C terminus to the trimerization domain of the bacteriophage T4 fibritin, termed ‘foldon’. The oligomerization status of recombinant RVG was investigated using sedimentation in sucrose gradient and p75^NTR binding assays. It was found that, in the absence of the fibritin foldon, recombinant RVGect forms unstable trimers that dissociate into monomers in a concentration-dependent manner. C-terminal fusion with the foldon induces stable RVG trimerization, which is concentration-independent. Furthermore, the fibritin foldon maintains the native antigenic structure of the carboxy part of RVGect. Cell binding experiments showed that RVG trimerization is required for efficient interaction with p75^NTR. However, the exact mode of trimerization appears unimportant, as trimeric recombinant RVGect (fused to the fibritin foldon) and RVGvir both recognize p75^NTR with similar nanomolar affinities, as shown by surface plasmon resonance experiments. Altogether, these results show that the C-terminal fusion of the RVG ectodomain with the fibritin foldon is a powerful way to obtain a recombinant trimeric native-like structure of the p75^NTR binding domain of RVG.

A supplementary table showing primers used in the construction of recombinant RVG clones is available as supplementary material in JGV Online.

These authors contributed equally to this work.

Present address: Laboratoire Immuno-Biotechnologie, Institut Pasteur de Tunis, 1002 Tunis-Belvédère, Tunisia.

This article has been cited by other articles:

				C. Tuffereau, K. Schmidt, C. Langevin, F. Lafay, G. Dechant, and M. Koltzenburg The Rabies Virus Glycoprotein Receptor p75NTR Is Not Essential for Rabies Virus Infection J. Virol., December 15, 2007; 81(24): 13622 - 13630. [Abstract] [Full Text] [PDF]

				B. Eschli, K. Quirin, A. Wepf, J. Weber, R. Zinkernagel, and H. Hengartner Identification of an N-Terminal Trimeric Coiled-Coil Core within Arenavirus Glycoprotein 2 Permits Assignment to Class I Viral Fusion Proteins. J. Virol., June 1, 2006; 80(12): 5897 - 5907. [Abstract] [Full Text] [PDF]