A linker strategy for the production and crystallization of Toll/interleukin-1 receptor/resistance protein domain complexes

Protein Engineering, Design & Selection : PEDS
Simon J WilliamsBostjan Kobe

Abstract

Structural characterization of protein-protein complexes is required to fully understand biological processes. However, such studies can be difficult, particularly when the interactions are transient. In some cases, the covalent linking of weakly interacting binding partners has been shown to facilitate structural studies. Here, we used this approach to investigate, by X-ray crystallography, the interactions between TIR (Toll/interleukin-1 receptor/resistance protein) domains from proteins involved in plant and animal innate immunity. Combinations of TIR domains known to interact were covalently attached using short glycine- and serine-rich linkers. This approach enabled the production of a number of TIR-TIR domain complexes in soluble form, facilitating crystallization studies. Crystallization of two of the tested combinations was achieved. Furthermore, production in soluble form was achieved for another two combinations, where this was not possible for individual proteins. Our results demonstrate that the linker strategy can aid in the structural studies of TIR domains. Similarly, this approach has potential for improving protein production and facilitating structural studies of other protein-protein interaction domains.

References

Dec 29, 2000·Nature·X XuX Wang
May 3, 2003·The EMBO Journal·Janet E DeaneJacqueline M Matthews
May 26, 2005·Protein Expression and Purification·F William Studier
May 16, 2006·Acta Crystallographica. Section D, Biological Crystallography·Sergei RadaevPeter D Sun
Jun 10, 2006·Science·Joseph D MougousJohn J Mekalanos
Nov 17, 2006·Nature·Jonathan D G Jones, Jeffery L Dangl
Jan 5, 2007·Annual Review of Immunology·Hyun Ho ParkHao Wu
Nov 22, 2008·Biochemical Society Transactions·Bostjan KobeJade K Forwood
Jan 10, 2009·Molecular Plant-microbe Interactions : MPMI·Michal R SwiderskiJonathan D G Jones
Jun 13, 2009·The Plant Journal : for Cell and Molecular Biology·Mari NarusakaYoshihiro Narusaka
Aug 19, 2009·The Plant Journal : for Cell and Molecular Biology·Doris BirkerRichard O'Connell
Aug 30, 2011·Proceedings of the National Academy of Sciences of the United States of America·Eugene ValkovBostjan Kobe
Feb 7, 2012·Current Opinion in Immunology·Vera BonardiJeffery L Dangl
Jun 6, 2012·Current Drug Targets·Thomas VeStuart Kellie
Dec 12, 2012·Protein Science : a Publication of the Protein Society·Vishnu Priyanka Reddy ChichiliJ Sivaraman
Aug 21, 2013·Science·Jeffery L DanglBrian J Staskawicz
Oct 9, 2013·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Li WanBostjan Kobe
Apr 20, 2014·Science·Simon J WilliamsJonathan D G Jones
Dec 3, 2014·Apoptosis : an International Journal on Programmed Cell Death·Thomas VeBostjan Kobe

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Citations

Jun 12, 2019·Experimental Biology and Medicine·Qianqian MingVincent C Luca
Jul 7, 2015·Acta Crystallographica. Section F, Structural Biology Communications·Bostjan KobeSimon J Williams

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