Crystallization and preliminary X-ray crystallographic analysis of importin-α from Neurospora crassa

• Published in: Acta crystallographica. Section F, Structural biology communications Structural biology communications, 2014-04, Vol.70 (4), p.501-504
• Main Authors: Bernardes, Natalia E., Takeda, Agnes A. S., Freitas, Fernanda Z., Bertolini, Maria Célia, Fontes, Marcos R. M.
• Format: Article
• Language: English
• Subjects: alpha Karyopherins - chemistry; alpha Karyopherins - genetics; alpha Karyopherins - metabolism; Cell Nucleus - metabolism; Crystallization - methods; Crystallization Communications; Crystallography, X-Ray - methods; importin-α; Neurospora crassa; Neurospora crassa - metabolism; NLS peptide; Oligopeptides - metabolism; Protein Binding; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism
• ISSN: 2053-230X; 2053-230X
• DOI: 10.1107/S2053230X14005068

Importin‐α recognizes cargo proteins that contain classical nuclear localization sequences (NLS) and, in complex with importin‐β, is able to translocate nuclear proteins through the nuclear pore complex. The filamentous fungus Neurospora crassa is a well studied organism that has been widely used as a model organism for fundamental aspects of eukaryotic biology, and is important for understanding the specific mechanisms of protein transport to the cell nucleus. In this work, the crystallization and preliminary X‐ray diffraction analysis of importin‐α from N. crassa (IMPα‐Nc) complexed with a classical NLS peptide (SV40 NLS) are reported. IMPα‐Nc–SV40 NLS crystals diffracted X‐rays to 2.0 Å resolution and the structure was solved by molecular‐replacement techniques, leading to a monomeric structure. The observation of the electron‐density map indicated the presence of SV40 NLSs interacting at both the minor and major NLS‐binding sites of the protein.