Design of an expression system to enhance MBP-mediated crystallization.

Sci Rep 2017 01 23;7:40991. Epub 2017 Jan 23.

Department of Pathology, Case Western Reserve University, Cleveland, OH 44106 USA.

Crystallization chaperones have been used to facilitate the crystallization of challenging proteins. Even though the maltose-binding protein (MBP) is one of the most commonly used crystallization chaperones, the design of optimal expression constructs for crystallization of MBP fusion proteins remains a challenge. To increase the success rate of MBP-facilitated crystallization, a series of expression vectors have been designed with either a short flexible linker or a set of rigid helical linkers. Seven death domain superfamily members were tested for crystallization with this set of vectors, six of which had never been crystallized before. All of the seven targets were crystallized, and their structures were determined using at least one of the vectors. Our successful crystallization of all of the targets demonstrates the validity of our approach and expands the arsenal of the crystallization chaperone toolkit, which may be applicable to crystallization of other difficult protein targets, as well as to other crystallization chaperones.

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http://www.nature.com/articles/srep40991
Publisher Site
http://dx.doi.org/10.1038/srep40991DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5256280PMC
January 2017
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