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【学术预告】Reactive Protein - Proteome Profiling “RP3”, a chemical approach to Fic-toxin substrate identification


报告题目:Reactive Protein - Proteome Profiling “RP3”, a chemical approach to Fic-toxin substrate identification

报告人:Prof. Dr. Christian Hedberg

工作单位:Institute of Chemistry, Ume? University, Sweden






Abstract:This presentation covers a new method denotedReactiveProtein -ProteomeProfilingRP3”.We have created a robust workflow for absolute substrate profiling of bacterial toxins against cellular target proteins employing covalent nucleotide co-substrates resulting in covalent ternary complexes. During infection, pathogenic bacteria tweak their eukaryotic hosts by translocating toxins. Toxins display catalytic activity against host cell proteins, targeting key functions, often using an abundant metabolite as co-substrate.No tools exist for the proteomic evaluation of the absolute substrate profile of a given toxin.Non-covalent affinity enrichment gives biased results depending on the reagent and relative target abundance. Our concept is based on covalent and subsequently cleavable capture of the target proteins by the reactive co-substrate of the toxin. Mutants (X to C) in the nucleotide-binding pocket of the toxin IbpA (H. somni)were modified with ATP-analogues carrying an electrophile. The binary complex forms the ternary complex with the IbpA-substrate cdc42in vitro. A number of other toxins belonging to the Fic-class have been profiled. For ADP-ribosylation, Pertussis toxin captures Giain vitrousing NAD+analogues. Altogether, this suggests that the RP3concept is valid and practicable.The project is highly significant for the infection biology field in terms of profiling toxins, but also to the chemical biology community, by moving ABPP to macromolecules via RP3.

报告人概况:Bengt Christian Hedberg博士,2005年获瑞典乌普萨拉必威官网有机化学博士。2008-2014担任德国马克斯普朗克分子生理学研究所化学生物学系Max-Planck研究组长,2014-至今担任瑞典于默奥必威官网化学研究所生物有机化学教授。2013年入选瑞典皇家科学院青年院士(Wallenberg Academy Fellow),近五年已在PNAS,Angewandte Chemie,Biophysical Journal,Journal of Organic Chemistry,Organic Letters等期刊发表论文15余篇。

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