Ketteler, R; Sun, ZR; Kovacs, KF; He, WW; Seed, B; (2008) A pathway sensor for genome-wide screens of intracellular proteolytic cleavage. GENOME BIOL , 9 (4) , Article R64. 10.1186/gb-2008-9-4-r64.
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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 beta-actin. By including protease cleavage sites between GLUC and beta-actin, proteolytic cleavage can be detected. Using this assay, we have identified regulators of autophagy, apoptosis and beta-actin cleavage.
|Title:||A pathway sensor for genome-wide screens of intracellular proteolytic cleavage|
|Keywords:||MAMMALIAN-CELLS, GAUSSIA LUCIFERASE, IN-VIVO, BIOLUMINESCENT REPORTER, SECRETED LUCIFERASE, PLASMA-MEMBRANE, GENE-EXPRESSION, CDNA CLONING, DEATH, AUTOPHAGY|
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