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제116회 대한화학회 학술발표회, 총회 및 기기전시회 안내 Unbiased Proteomic Profiling Strategy for Discovery of Bacterial Effector Proteins Reveals that Salmonella Protein PheA Is a Host Cell Cycle Regulator

2015년 9월 3일 14시 42분 23초
BIO.P-217 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
10월 15일 (목요일) 11:00~12:30
저자 및
나하나, 이준석1,*
과학기술연합대학원대학교(UST) 생물화학, Korea
1한국과학기술연구원(KIST) 분자인지연구센터, Korea
Salmonella can survive inside macrophages through injecting effector proteins into the host cell cytosol and cause various diseases. It is not easy to identify effector protein because of low throughput issue of GFP tagging-based genetic manipulation. Herein, we demonstrate that off-the-shelf fluorescent probes, especially, Flu-NHS that contain N-hydroxysuccinimide functional groups with a negatively charge state can tag salmonella proteins including effector protein in RAW264.7 macrophage cells infected with salmonella in the unbiased way. We use competitive chemical proteomic profiling method and bioinformatics analysis such as cross-species BLAST analysis, sequence analysis and draw one top among 29 potential candidates. That is chorismate mutase-P/prehenate dehydrates, PheA. PheA is a Salmonella enzyme known for phenyl alanine biosynthesis and metabolic processing of chorismate. This protein exhibited significant sequence similarity to an area near the DNA binding domain of E2F transcription factor 7, E2F7. The functional similarity such as cell cycle alteration promoting G1/S cell cycle arrest is discovered using FACS and electrophoretic mobility shift assay. Based on our results, we have demonstrated the power of an unbiased tagging approach for the discovery of host cell regulators from pathogens.