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Application of linker technique to trap transiently interacting protein complexes for structural studies.

J Biol Methods. 2016; 
Reddy Chichili VP, Kumar V, Sivaraman J.
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摘要

Protein-protein interactions are key events controlling several biological processes. We have developed and employed a method to trap transiently interacting protein complexes for structural studies using glycine-rich linkers to fuse interacting partners, one of which is unstructured. Initial steps involve isothermal titration calorimetry to identify the minimum binding region of the unstructured protein in its interaction with its stable binding partner. This is followed by computational analysis to identify the approximate site of the interaction and to design an appropriate linker length. Subsequently, fused constructs are generated and characterized using size exclusion chromatography and dynamic light scat... More

关键词

glycine-rich linker; protein-protein interactions; transient interactions