EngineeringPhotorhabdus luminescenstoxin complex (PTC) into a recombinant injection nanomachine

Author:

Ng´ang´a Peter Njenga123,Ebner Julia K123,Plessner Matthias1,Aktories Klaus14,Schmidt Gudula1ORCID

Affiliation:

1. Institute for Experimental and Clinical Pharmacology and Toxicology, Faculty of Medicine, University of Freiburg, Freiburg, Germany

2. Faculty of Biology, University of Freiburg, Freiburg, Germany

3. Spemann Graduate School for Biology and Medicine, University of Freiburg, Freiburg, Germany

4. BIOSS Centre for Biological Signalling Studies, University of Freiburg, Freiburg, Germany

Abstract

Engineering delivery systems for proteins and peptides into mammalian cells is an ongoing challenge for cell biological studies as well as for therapeutic approaches.Photorhabdus luminescenstoxin complex (PTC) is a heterotrimeric protein complex able to deliver diverse protein toxins into mammalian cells. We engineered the syringe-like nanomachine for delivery of protein toxins from different species. In addition, we loaded the highly active copepod luciferaseMetridia longaM-Luc7 for accurate quantification of injected molecules. We suggest that besides the probable size limitation, the charge of the cargo also influences the efficiency of packing and transport into mammalian cells. Our data show that the PTC constitutes a powerful system to inject recombinant proteins, peptides, and potentially, other molecules into mammalian cells. In addition, in contrast to other protein transporters based on pore formation, the closed, compact structure of the PTC may protect cargo from degradation.

Funder

Deutsche Forschungsgemeinschaft SFB 850

DAAD

Publisher

Life Science Alliance, LLC

Subject

Health, Toxicology and Mutagenesis,Plant Science,Biochemistry, Genetics and Molecular Biology (miscellaneous),Ecology

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