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Open Access Methodology article

Application of Gaussia luciferase in bicistronic and non-conventional secretion reporter constructs

Christin Luft1, Jamie Freeman1, David Elliott1, Nadia Al-Tamimi1, Janos Kriston-Vizi1, Jacob Heintze1, Ida Lindenschmidt1, Brian Seed2 and Robin Ketteler1*

Author Affiliations

1 Medical Research Council, Laboratory for Moleclar and Cell Biology, University College London, Gower Street, London WC1E 6BT, UK

2 Massachusetts General Hospital, Center for Computational and Integrative Biology, Boston MA02114, USA

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BMC Biochemistry 2014, 15:14  doi:10.1186/1471-2091-15-14

Published: 9 July 2014

Abstract

Background

Secreted luciferases are highly useful bioluminescent reporters for cell-based assays and drug discovery. A variety of secreted luciferases from marine organisms have been described that harbor an N-terminal signal peptide for release along the classical secretory pathway. Here, we have characterized the secretion of Gaussia luciferase in more detail.

Results

We describe three basic mechanisms by which GLUC can be released from cells: first, classical secretion by virtue of the N-terminal signal peptide; second, internal signal peptide-mediated secretion and third, non-conventional secretion in the absence of an N-terminal signal peptide. Non-conventional release of dNGLUC is not stress-induced, does not require autophagy and can be enhanced by growth factor stimulation. Furthermore, we have identified the golgi-associated, gamma adaptin ear containing, ARF binding protein 1 (GGA1) as a suppressor of release of dNGLUC.

Conclusions

Due to its secretion via multiple secretion pathways GLUC can find multiple applications as a research tool to study classical and non-conventional secretion. As GLUC can also be released from a reporter construct by internal signal peptide-mediated secretion it can be incorporated in a novel bicistronic secretion system.

Keywords:
Non-conventional secretion; Gaussia luciferase; GGA1; Bicistronic expression; Signal peptide