Construct optimization for studying protein complexes: obtaining diffraction‐quality crystals of the pseudosymmetric PSPC1–NONO heterodimer
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
Publication date: November 1, 2011