This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Segura, M. M.
Right arrow Articles by Kamen, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Segura, M. M.
Right arrow Articles by Kamen, A.

 Previous Article  |  Next Article 

Journal of Virology, February 2008, p. 1107-1117, Vol. 82, No. 3
0022-538X/08/$08.00+0     doi:10.1128/JVI.01909-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Identification of Host Proteins Associated with Retroviral Vector Particles by Proteomic Analysis of Highly Purified Vector Preparations{triangledown}

María Mercedes Segura,1,2,{dagger} Alain Garnier,2 Marcos Rafael Di Falco,3 Gavin Whissell,1,{ddagger} Angélica Meneses-Acosta,1,§ Normand Arcand,1 and Amine Kamen1*

Biotechnology Research Institute, NRC, 6100 Royalmount Avenue, Montreal, Quebec, Canada H4P 2R2,1 Department of Chemical Engineering, Centre de Recherche sur la fonction, la structure et l'ingénierie des protéines, Université Laval, Québec, Canada G1K 7P4,2 Genome Quebec Innovation Centre, McGill University, 740 Dr Penfield, Montreal, Quebec, Canada H3A 1A43

Received 31 August 2007/ Accepted 5 November 2007

The Moloney murine leukemia virus (MMLV) belongs to the Retroviridae family of enveloped viruses, which is known to acquire minute amounts of host cellular proteins both on the surface and inside the virion. Despite the extensive use of retroviral vectors in experimental and clinical applications, the repertoire of host proteins incorporated into MMLV vector particles remains unexplored. We report here the identification of host proteins from highly purified retroviral vector preparations obtained by rate-zonal ultracentrifugation. Viral proteins were fractionated by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis, in-gel tryptic digested, and subjected to liquid chromatography/tandem mass spectrometry analysis. Immunogold electron microscopy studies confirmed the presence of several host membrane proteins exposed at the vector surface. These studies led to the identification of 27 host proteins on MMLV vector particles derived from 293 HEK cells, including 5 proteins previously described as part of wild-type MMLV. Nineteen host proteins identified corresponded to intracellular proteins. A total of eight host membrane proteins were identified, including cell adhesion proteins integrin β1 (fibronectin receptor subunit beta) and HMFG-E8, tetraspanins CD81 and CD9, and late endosomal markers CD63 and Lamp-2. Identification of membrane proteins on the retroviral surface is particularly attractive, since they can serve as anchoring sites for the insertion of tags for targeting or purification purposes. The implications of our findings for retrovirus-mediated gene therapy are discussed.

* Corresponding author. Mailing address: Biotechnology Research Institute, NRC, 6100 Royalmount Avenue, Montreal, Quebec, Canada H4P 2R2. Phone: (514) 496-2264. Fax: (514) 496-7251. E-mail: amine.kamen{at}cnrc-nrc.gc.ca

{triangledown} Published ahead of print on 21 November 2007.

{dagger} Present address: Center of Animal Biology and Gene Therapy (CBATEG), Universitat Autònoma de Barcelona, Bellaterra (08193), Barcelona, Spain.

{ddagger} Present address: Institute for Research in Biomedicine (IRB), Parc Científic de Barcelona, C/Josep Samitier 1-5 (08028), Barcelona, Spain.

§ Present address: Facultad de Farmacia, Universidad Autónoma del Estado de Morelos, Cuernavaca (62210), Morelos, México.

Journal of Virology, February 2008, p. 1107-1117, Vol. 82, No. 3
0022-538X/08/$08.00+0     doi:10.1128/JVI.01909-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Tan, Y., Spears, T., Bideshi, D. K., Johnson, J. J., Hice, R., Bigot, Y., Federici, B. A. (2009). P64, a Novel Major Virion DNA-Binding Protein Potentially Involved in Condensing the Spodoptera frugiperda Ascovirus 1a Genome. J. Virol. 83: 2708-2714 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»