Engineering an AB₅ Protein Carrier

Title data

Lichtenstein, Bruce R. ; Höcker, Birte:
Engineering an AB₅ Protein Carrier.
In: Scientific Reports. Vol. 8 (2018) . - 12643.
ISSN 2045-2322
DOI: https://doi.org/10.1038/s41598-018-30910-y

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Abstract in another language

The promise of biologic therapeutics is hindered by the challenge to deliver their activity to biochemically relevant sites within diseased cells. The favourable application of the natural protein carriers of the AB₅ toxin family to this challenge has been restricted owing to still unresolved requirements for assembling non-native cargo into carrier complexes. Here, we clarify the properties of fusion peptides which allow co-assembly of a selected fluorescent protein cargo with the non-toxic B subunit of a heat-labile enterotoxin. We establish the influence of sequence length, sequence identity and secondary structure of these linking domains on the assembly and disassembly of the complexes. Through our engineering framework we identify several non-native, reduced length fusion sequences that robustly assemble with the native carriers, maintain their ability to deliver protein cargo to cells, and demonstrate substantially refined in vitro properties. Constructs based upon these sequences should prove directly applicable to a variety of protein delivery challenges, and the described design framework should find immediate application to other members of the AB₅ protein carrier family.