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Construct optimization for studying protein complexes: obtaining diffraction‐quality crystals of the pseudosymmetric PSPC1–NONO heterodimer

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Abstract:

The methodology of protein crystallography provides a number of potential bottlenecks. Here, an approach to successful structure solution of a difficult heterodimeric complex of two human proteins, paraspeckle component 1 (PSPC1) and non‐POU domain‐containing octamer‐binding protein (NONO), that are involved in gene regulation and the¬†structural integrity of nuclear bodies termed paraspeckles is¬†described. With the aid of bioinformatic predictions and systematic screening of a panel of constructs, bottlenecks of protein solubility, crystallization, crystal quality and crystallographic pseudosymmetry were overcome in order to produce crystals that ultimately revealed the structure.

Document Type: Research Article

DOI: http://dx.doi.org/10.1107/S0907444911039606

Publication date: November 1, 2011

bsc/ayd/2011/00000067/00000011/art00010
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