Purification and crystallization of human Cu/Zn superoxide dismutase recombinantly produced in the protozoan Leishmania tarentolae

0347295 - BC 2011 RIV GB eng J - Článek v odborném periodiku
Gazdag, E. M. - Cirstea, I. - Breitling, R. - Lukeš, Julius - Blankenfeldt, W. - Alexandrov, K.
Purification and crystallization of human Cu/Zn superoxide dismutase recombinantly produced in the protozoan Leishmania tarentolae.
Acta Crystallographica Section F-Structural Biology and Crystallization Communications. Roč. 66, č. 8 (2010), s. 871-877. ISSN 1744-3091. E-ISSN 2053-230X
Výzkumný záměr: CEZ:AV0Z60220518
Klíčová slova: recombinant proteins * eukaryotic expression systems * Leishmania
Kód oboru RIV: EB - Genetika a molekulární biologie
Impakt faktor: 0.563, rok: 2010

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.
Trvalý link: http://hdl.handle.net/11104/0188101