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A highly sensitive protein-protein interaction assay based on Gaussia luciferase


Protein-fragment complementation assays (PCAs) provide a general strategy to study the dynamics of protein-protein interactions in vivo and in vitro. The full potential of PCA requires assays that are fully reversible and sensitive at subendogenous protein expression levels. We describe a new assay that meets these criteria, based on the Gaussia princeps luciferase enzyme, demonstrating chemical reversal, and induction and inhibition of a key interaction linking insulin and TGFβ signaling.

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Figure 1: Characterization of the hGLuc PCA.
Figure 2: The hGLuc PCA is inducible and fully reversible.
Figure 3: Endogenous levels of interacting proteins can be detected with the hGLuc PCA.


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We gratefully acknowledge V. Rivera and T. Clackson (ARIAD Pharmaceuticals, Inc.) for providing AP21998 and FM cDNA. This work was supported by the Canadian Institute of Health Research (MOP-152556).

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Correspondence to Stephen W Michnick.

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S.W.M. is a common stock holder in Odyssey Thera Inc.

Supplementary information

Supplementary Fig. 1

Development of the Gaussia luciferase PCA. (PDF 283 kb)

Supplementary Fig. 2

Reassociation of FM-hGLuc(1)/FM-hGLuc(2) complexes after AP21998 removal. (PDF 112 kb)

Supplementary Fig. 3

Disruption of FM-hGLuc(1)/FM-hGLuc(2) complexes by FK506. (PDF 135 kb)

Supplementary Fig. 4

hGLuc and hRLuc PCAs fold within 60 seconds. (PDF 162 kb)

Supplementary Methods (PDF 87 kb)

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Remy, I., Michnick, S. A highly sensitive protein-protein interaction assay based on Gaussia luciferase. Nat Methods 3, 977–979 (2006).

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