Email updates

Keep up to date with the latest news and content from Genome Biology and BioMed Central.

Open Access Highly Accessed Method

A pathway sensor for genome-wide screens of intracellular proteolytic cleavage

Robin Ketteler12, Zairen Sun3, Karl F Kovacs3, Wei-Wu He3 and Brian Seed12*

Author Affiliations

1 Center for Computational and Integrative Biology, Massachusetts General Hospital, Cambridge Street, Boston, MA 02114, USA

2 Department of Genetics, Harvard Medical School, Cambridge Street, Boston, MA 02114, USA

3 Origene Technologies Inc., Taft Court, Rockville, MD 20850, USA

For all author emails, please log on.

Genome Biology 2008, 9:R64  doi:10.1186/gb-2008-9-4-r64

Published: 3 April 2008

Abstract

Protein cleavage is a central event in many regulated biological processes. We describe a system for detecting intracellular proteolysis based on non-conventional secretion of Gaussia luciferase (GLUC). GLUC exits the cell without benefit of a secretory leader peptide, but can be anchored in the cell by fusion to β-actin. By including protease cleavage sites between GLUC and β-actin, proteolytic cleavage can be detected. Using this assay, we have identified regulators of autophagy, apoptosis and β-actin cleavage.