title: Purification and crystallization of human Cu/Zn superoxide dismutase recombinantly produced in the protozoan Leishmania tarentolae.
abstract: The rapid and inexpensive production of high-quality eukaryotic proteins in recombinant form still remains a challenge in structural biology. Here, a protein-expression system based on the protozoan Leishmania tarentolae was used to produce human Cu/Zn superoxide dismutase (SOD1) in recombinant form. Sequential integration of the SOD1 expression cassettes was demonstrated to lead to a linear increase in expression levels to up to 30 mg per litre. Chromatographic purification resulted in 90% pure recombinant protein, with a final yield of 6.5 mg per litre of culture. The protein was crystallized and the structures of two new crystal forms were determined. These results demonstrate the suitability of the L. tarentolae expression system for structural research.
authors.affiliation: Department of Physical Biochemistry, Max-Planck-Institute for Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund, Germany.
authors.forename: Emerich Mihai
authors.initials: EM
authors.lastname: Gazdag
authors.forename: Ion Cristian
authors.initials: IC
authors.lastname: Cirstea
authors.forename: Reinhard
authors.initials: R
authors.lastname: Breitling
authors.forename: Julius
authors.initials: J
authors.lastname: Lukes
authors.forename: Wulf
authors.initials: W
authors.lastname: Blankenfeldt
authors.forename: Kirill
authors.initials: K
authors.lastname: Alexandrov
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D006801:Humans; D007891:Leishmania / Q000378:metabolism*; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D013482:Superoxide Dismutase