research papers
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.
Keywords: PSPC1-NONO heterodimer; paraspeckles; construct design; pseudosymmetry; RRM; coiled coils.
Supporting information
Portable Document Format (PDF) file https://doi.org/10.1107/S0907444911039606/gx5190sup1.pdf |