Engineered bacterial outer membrane vesicles with enhanced functionality

JOURNAL OF MOLECULAR BIOLOGY

卷:380

Article

Kim, Jae-Young¹  Doody, Anne M.^1,2  Chen, David J.¹  Cremona, Gina H.¹  Shuler, Michael L.^1,2  Putnam, David^1,2  DeLisa, Matthew P.^1,2 

[1] Cornell Univ, Sch Chem & Biomol Engn, Ithaca, NY 14853 USA

[2] Cornell Univ, Dept Biomed Engn, Ithaca, NY 14853 USA

关键词: blebs;    cytotoxin;    protein secretion;    vaccines;    virulence factors;   

DOI  :  10.1016/j.jmb.2008.03.076

来源: Elsevier

【 摘 要 】

We have engineered bacterial outer membrane vesicles (OMVs) with dramatically enhanced functionality by fusing several heterologous proteins to the vesicle-associated toxin ClyA of Escherichia coli. Similar to native unfused ClyA, chimeric ClyA fusion proteins were found localized in bacterial OMVs and retained activity of the fusion partners, demonstrating for the first time that ClyA can be used to co-localize fully functional heterologous proteins h e enzymes directly in bacterial OMVs. For instance, fusions of ClyA to the enzymes beta-lactamase and organophosphorus hydrolase resulted in synthetic OMVs that were capable of hydrolyzing beta-lactam antibiotics and paraoxon, respectively. Similarly, expression of an anti-digoxin single-chain Fv antibody fragment fused to the C terminus of ClyA resulted in designer immuno-MVs that could bind tightly and specifically to the antibody's cognate antigen. Finally, OMVs displaying green fluorescent protein fused to the C terminus of ClyA were highly fluorescent and, as a result of this new functionality, could be easily tracked during vesicle interaction with human epithelial cells. We expect that the relative plasticity exhibited by ClyA as a fusion partner should prove useful for: (i) further mechanistic studies to identify the vesiculation machinery that regulates OMV secretion and to map the intracellular routing of ClyA-containing OMVs during invasion of host cells; and (ii) biotechnology applications such as surface display of proteins and delivery of biologics. (C) 2008 Elsevier Ltd. All rights reserved.

【 授权许可】

Free