Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học Critical Care giúp cho các bạn có thêm kiến thức về ngành y học đề tài: A pathway sensor for genome-wide screens of intracellular proteolytic cleavage. | Open Access Method A pathway sensor for genome-wide screens of intracellular proteolytic cleavage Robin Ketteler Zairen Sun Karl F Kovacs Wei-Wu He and Brian Seed Addresses Center for Computational and Integrative Biology Massachusetts General Hospital Cambridge Street Boston MA 02114 USA. Department of Genetics Harvard Medical School Cambridge Street Boston MA 02114 USA. Origene Technologies Inc. Taft Court Rockville MD 20850 USA. Correspondence Brian Seed. Email bseed@ Published 3 April 2008 Genome Biology 2008 9 R64 doi 186 gb-2008-9-4-r64 The electronic version of this article is the complete one and can be found online at http 2008 9 4 R64 Received 25 February 2008 Revised 19 March 2008 Accepted 3 April 2008 2008 Ketteler et al. licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License http licenses by which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. 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 P-actin. By including protease cleavage sites between GLUC and P-actin proteolytic cleavage can be detected. Using this assay we have identified regulators of autophagy apoptosis and P-actin cleavage. Background Advances in automation and the availability of genomic sequence information have led to the development of sophisticated cell-based assays for high-throughput screening of functional phenotypes 1 . Most cell-based assays rely on fluorescent or luminescent reporters such as green fluorescent protein GFP secreted alkaline phosphatase SEAP or Photinus luciferase. Secreted luciferases
