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Chemoselective DNA conjugation of proteins to study multienzyme clomplexes

Chemoselective DNA conjugation of proteins to study multienzyme clomplexes
蛋白质的化学选择性 DNA 缀合研究多酶复合物
批准号:
223269940
负责人:
Professor Dr. Christof M. Niemeyer
金额:
$0.0万
依托单位国家:
德国
项目类别:
Priority Programmes
财政年份:
2012
资助国家:
德国
项目状态:
已结题
起止时间:
2011-12-31 至 2015-12-31

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中文摘要
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英文摘要
This project concerns the development of a generic platform to generate multfunctional protein constructs useful for interrogation of biological systems. The platform technology is based on the combination of chemoselective protein ligation methods and structural DNA nanotechnology. In particular, we will establish the chemoenzymatic ligation of recombinant proteins derived from multimodular Polyketide Synthases (PKS) with DNA or PNA oligonucleotides, which are derivatized with a short peptide tag, serving as the ligation substrate of bacterial transpeptidase Sortase. The resulting DNA-/PNA-tagged PKS modules will be assembled by Watson-Crick base pairing in homogeneous solution and, in particular, on the surface of two-dimensional nucleic acid scaffolds generated by the DNA origami technique. In a highly modular approach, supramolecular multienzyme complexes will be built, in which the spatial configuration of the PKS domains is controlled by the DNA scaffold with nanometer precision. On the one hand, this project will contribute to the detailed understanding of the spatial organization, conformational flexibility and other fundamental principles governing multienzyme complexes. On the other hand, the platform developed here will enable ready access to numerous spatially well-defined multifunctional protein complexes for mimicking, manipulation and analysis of natural occuring supramolecular assemblies involved in gene regulation, signal transduction, division, or other cellular function and traits.
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