Quantitative proteomic analysis of lentiviral vectors using 2-DE
| Autor: | Florence Gonnet, Olivier Danos, Otto Wilhelm Merten, Marina Radrizzani, Luigi Naldini, Anne Galy, Jérôme Denard, Nicolas Laroudie, Fedor Svinartchouk, Stéphanie Rundwasser |
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| Přispěvatelé: | Denard, J, Rundwasser, S, Laroudie, N, Gonnet, F, Naldini, Luigi, Radrizzani, M, Galy, A, Merten, Ow, Danos, O, Svinartchouk, F. |
| Rok vydání: | 2009 |
| Předmět: |
Proteomics
Genetic enhancement Genetic Vectors Lentivirus Proteins Biology Vectors in gene therapy biology.organism_classification Biochemistry Virology Virus Cell Line Viral vector law.invention Viral Proteins law Vesicular stomatitis virus Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Recombinant DNA Humans Electrophoresis Gel Two-Dimensional Vector (molecular biology) Molecular Biology |
| Popis: | Among the integrative gene therapy vectors developed to date, human immunodeficiency virus type 1 (HIV-1)-derived lentiviral vectors (LV) are distinguished by their capacity to infect both dividing and non-dividing cells. Recombinant LV particles contain viral proteins necessary for their packaging, infectious and integrating functions. Like the parental HIV-1 virus they are able to acquire various cellular proteins, but the number and localisation of these proteins are poorly characterised. In the present study we used 2-DE followed by MALDI-TOF to quantify the protein content of several types of vesicular stomatitis virus G-pseudotyped LV including those that were extensively purified in the perspective of clinical gene therapy studies. A proteinase K treatment was used to distinguish between cellular proteins incorporated into virions (I-proteins) and those co-purified with vectors (C-proteins). We found 10 C-proteins and 18 I-proteins associated with LV. Copy numbers for these core I-proteins varied from 5 (AIP-1/ALIX) to 280 (Cyclophilin A) per vector particle. Three novel I-proteins, guanine nucleotide-binding protein 2, L-lactate dehydrogenase B chain and hnRNP core protein A1, were found. This study defines for the first time, the protein stoichiometry of infectious HIV-1-derived LV particles. |
| Databáze: | OpenAIRE |
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