3KH3

Crystal structure of human Cu/Zn superoxide dismutase recombinantly produced in Leishmania tarantolae; P212121 crystal form containing 12 chains in the asymmetric unit

Summary for 3KH3

• Entry DOI: 10.2210/pdb3kh3/pdb
• Related: 3KH4
• Descriptor: Superoxide dismutase [Cu-Zn], COPPER (II) ION, ZINC ION, ... (4 entities in total)
• Functional Keywords: eukaryotic expression, leishmania tarantolae, amyotrophic lateral sclerosis, antioxidant, disease mutation, disulfide bond, metal-binding, neurodegeneration, oxidoreductase, phosphoprotein
• Biological source: Homo sapiens (human)
• Cellular location: Cytoplasm: P00441
• Total number of polymer chains: 12
• Total formula weight: 191766.38

Authors

Gazdag, E.M.,Blankenfeldt, W. (deposition date: 2009-10-30, release date: 2010-08-11, Last modification date: 2024-10-16)

Primary citation

Gazdag, E.M.,Cirstea, I.C.,Breitling, R.,Lukes, J.,Blankenfeldt, W.,Alexandrov, K.
Purification and crystallization of human Cu/Zn superoxide dismutase recombinantly produced in the protozoan Leishmania tarentolae.
Acta Crystallogr.,Sect.F, 66:871-877, 2010

PubMed 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.

PubMed: 20693657
DOI: 10.1107/S1744309110019330

Experimental method

X-RAY DIFFRACTION (3.5 Å)