Number of the records: 1
Purification and crystallization of human Cu/Zn superoxide dismutase recombinantly produced in the protozoan Leishmania tarentolae
- 1. 0347295 - BC-A 2011 RIV GB eng J - Journal Article
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. Roč. 66, č. 8 (2010), s. 871-877. ISSN 1744-3091
Institutional research plan: CEZ:AV0Z60220518
Keywords : recombinant proteins * eukaryotic expression systems * Leishmania
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 0.563, year: 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.
Permanent Link: http://hdl.handle.net/11104/0188101