Format

Send to

Choose Destination
Acta Crystallogr F Struct Biol Commun. 2015 Dec;71(Pt 12):1481-7. doi: 10.1107/S2053230X15021524. Epub 2015 Nov 18.

Combining dehydration, construct optimization and improved data collection to solve the crystal structure of a CRM1-RanGTP-SPN1-Nup214 quaternary nuclear export complex.

Author information

1
Abteilung für Molekulare Strukturbiologie, Institut für Mikrobiologie und Genetik, Göttinger Zentrum für Molekulare Biowissenschaften (GZMB), Georg-August-Universität Göttingen, Justus-von-Liebig-Weg 11, 37077 Göttingen, Germany.
2
Macromolecular Crystallography (HZB-MX), Helmholtz-Zentrum Berlin für Materialien und Energie, Albert-Einstein-Strasse 15, 12489 Berlin, Germany.
3
Institut für Molekularbiologie, Universitätsmedizin Göttingen, Georg-August-Universität Göttingen, Humboldtallee 23, 37073 Göttingen, Germany.

Abstract

High conformational flexibility is an intrinsic and indispensable property of nuclear transport receptors, which makes crystallization and structure determination of macromolecular complexes containing exportins or importins particularly challenging. Here, the crystallization and structure determination of a quaternary nuclear export complex consisting of the exportin CRM1, the small GTPase Ran in its GTP-bound form, the export cargo SPN1 and an FG repeat-containing fragment of the nuclear pore complex component nucleoporin Nup214 fused to maltose-binding protein is reported. Optimization of constructs, seeding and the development of a sophisticated protocol including successive PEG-mediated crystal dehydration as well as additional post-mounting steps were essential to obtain well diffracting crystals.

KEYWORDS:

CRM1; HC1c crystal humidifier; crystal dehydration; maltose-binding protein; nucleoporin

PMID:
26625290
PMCID:
PMC4666476
DOI:
10.1107/S2053230X15021524
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for International Union of Crystallography Icon for PubMed Central
Loading ...
Support Center