TY - JOUR
T1 - Construct optimization for studying protein complexes: obtaining diffraction-quality crystals of the pseudosymmetric PSPC1-NONO heterodimer
AU - Lee, Mihwa
AU - Passon, Daniel M.
AU - Hennig, Sven
AU - Fox, Archa H.
AU - Bond, Charles S.
PY - 2011/11/9
Y1 - 2011/11/9
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/80955123973
U2 - 10.1107/S0907444911039606
DO - 10.1107/S0907444911039606
M3 - Article
SN - 0907-4449
VL - 67
SP - 981
EP - 987
JO - Acta Crystallographica. Section D: Biological Crystallography
JF - Acta Crystallographica. Section D: Biological Crystallography
IS - 11
ER -