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

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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.
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Document Type: Research Article

Publication date: 2011-11-01

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