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Comparison of Membrane Targeting Strategies for the Accumulation of the Human Immunodeficiency Virus p24 Protein in Transgenic Tobacco

1
Department of Plant Cell Biology, Centre for Organismal Studies, University of Heidelberg, Heidelberg D-69120, Germany
2
Institute of Agricultural Biology and Biotechnology, National Research Council (CNR), via Bassini 15, Milano 20133, Italy
3
Laboratory of Cell and Molecular Biology, University of Neuchatel, Rue Emile-Argand 11, Neuchâtel CH-2000, Switzerland
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2013, 14(7), 13241-13265; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms140713241
Received: 7 April 2013 / Revised: 28 May 2013 / Accepted: 17 June 2013 / Published: 26 June 2013
(This article belongs to the Section Biochemistry)
Membrane anchorage was tested as a strategy to accumulate recombinant proteins in transgenic plants. Transmembrane domains of different lengths and topology were fused to the cytosolic HIV antigen p24, to promote endoplasmic reticulum (ER) residence or traffic to distal compartments of the secretory pathway in transgenic tobacco. Fusions to a domain of the maize seed storage protein γ-zein were also expressed, as a reference strategy that leads to very high stability via the formation of large polymers in the ER lumen. Although all the membrane anchored constructs were less stable compared to the zein fusions, residence at the ER membrane either as a type I fusion (where the p24 sequence is luminal) or a tail-anchored fusion (where the p24 sequence is cytosolic) resulted in much higher stability than delivery to the plasma membrane or intermediate traffic compartments. Delivery to the tonoplast was never observed. The inclusion of a thrombin cleavage site allowed for the quantitative in vitro recovery of p24 from all constructs. These results point to the ER as suitable compartment for the accumulation of membrane-anchored recombinant proteins in plants. View Full-Text
Keywords: HIV p24; transgenic plants; membrane proteins; endomembrane system; protein targeting; thrombin cleavage HIV p24; transgenic plants; membrane proteins; endomembrane system; protein targeting; thrombin cleavage
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MDPI and ACS Style

Virgili-López, G.; Langhans, M.; Bubeck, J.; Pedrazzini, E.; Gouzerh, G.; Neuhaus, J.-M.; Robinson, D.G.; Vitale, A. Comparison of Membrane Targeting Strategies for the Accumulation of the Human Immunodeficiency Virus p24 Protein in Transgenic Tobacco. Int. J. Mol. Sci. 2013, 14, 13241-13265. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms140713241

AMA Style

Virgili-López G, Langhans M, Bubeck J, Pedrazzini E, Gouzerh G, Neuhaus J-M, Robinson DG, Vitale A. Comparison of Membrane Targeting Strategies for the Accumulation of the Human Immunodeficiency Virus p24 Protein in Transgenic Tobacco. International Journal of Molecular Sciences. 2013; 14(7):13241-13265. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms140713241

Chicago/Turabian Style

Virgili-López, Goretti, Markus Langhans, Julia Bubeck, Emanuela Pedrazzini, Guillaume Gouzerh, Jean-Marc Neuhaus, David G. Robinson, and Alessandro Vitale. 2013. "Comparison of Membrane Targeting Strategies for the Accumulation of the Human Immunodeficiency Virus p24 Protein in Transgenic Tobacco" International Journal of Molecular Sciences 14, no. 7: 13241-13265. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms140713241

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