Crystallization and preliminary X-ray crystallographic analysis of importin-α from Neurospora crassa.
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.
Departamento de Física e Biofísica, Instituto de Biociências, UNESP - Universidade Estadual Paulista, Botucatu, SP, Brazil.
Natalia E
NE
Bernardes
Departamento de Física e Biofísica, Instituto de Biociências, UNESP - Universidade Estadual Paulista, Botucatu, SP, Brazil.
Agnes A S
AA
Takeda
Departamento de Bioquímica e Tecnologia Química, Instituto de Química, UNESP - Universidade Estadual Paulista, Araraquara, SP, Brazil.
Fernanda Z
FZ
Freitas
Departamento de Bioquímica e Tecnologia Química, Instituto de Química, UNESP - Universidade Estadual Paulista, Araraquara, SP, Brazil.
Maria Célia
MC
Bertolini
Departamento de Física e Biofísica, Instituto de Biociências, UNESP - Universidade Estadual Paulista, Botucatu, SP, Brazil.
Marcos R M
MR
Fontes
D002467:Cell Nucleus / Q000378:metabolism; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray / Q000379:methods*; D009492:Neurospora crassa / Q000378:metabolism*; D009842:Oligopeptides / Q000378:metabolism*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D028901:alpha Karyopherins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D009842:Oligopeptides; D011994:Recombinant Proteins; C402304:SV40 nuclear localization peptide; D028901:alpha Karyopherins
NLS peptide; Neurospora crassa; importin-α