A Simple Tagging System for Protein Encapsulation

A Simple Tagging System for Protein Encapsulation
Received December 9, 2005
Web Release Date: March 22, 2006
Florian P. Seebeck, Kenneth J. Woycechowsky, Wei Zhuang, J?rgen P. Rabe, and Donald Hilvert*
J. Am. Chem. Soc.
ACS Publications
Copyright ? 2006 American Chemical Society
Laboratorium f?r Organische Chemie, ETH Z?rich, H?nggerberg HCI F337, CH-8093, Z?rich, Switzerland, and Department of Physics, Humboldt University Berlin, Newtonstrasse 15, D-12489 Berlin, Germany
Molecular containers that encapsulate specific cargo can be useful for many natural and non-natural processes. We report a simple system, based on charge complementarity, for the encapsulation of appropriately tagged proteins within an engineered, proteinaceous capsid. Four negative charges per monomer were added to the lumazine synthase from Aquifex aeolicus (AaLS). The capsids formed by the engineered AaLS associate with green fluorescent protein bearing a positively charged deca-arginine tag upon coproduction in Escherichia coli. Analytical ultracentrifugation and scanning force microscopy studies indicated that the engineered AaLS retains the ability to form capsids, but that their average size was substantially increased. The success of this strategy demonstrates that both the container and guest components of protein-based encapsulation systems can be convergently designed in a straightforward manner, which may help to extend their versatility.
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