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. 2017 Aug 15;7:8186. doi: 10.1038/s41598-017-07569-y

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Design of a dual peptide complementation system. (A) Topological representation of the 10 β-stranded structure of NanoLuc (adapted from Dixon et al.¹⁶ with permission from ACS Chem. Bio.). The ternary system consists of strands 1–8 (Δ11S) and two 11 amino acid peptides stemming from strands 9 (β9, red) and 10 (β10, blue). Sequences of Δ11S, β9, and β10 are listed in supplemental information, Table S1. (B) Structural model illustrating the activation of Δ11S only in the presence of both β9 and β10. Model was created from PDB ID 5IBO. (C) Luminescence produced from different combinations of the complementation components. Luminescence is only produced with all three components, ***p < 0.001. Concentrations of β9, β10, and Δ11S were 1 μM, 1 μM and 1:10⁵ dilution of lysate, respectively. (D) The luminescent signal over time following the addition of Δ11S (1:10⁵ dilution of lysate) to a solution of 1 μM β9 and β10. Under these conditions, full complementation occurs in under 20 minutes.