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Expression, purification and crystallization of Chaetomium thermophilum Cu,Zn superoxide dismutase.

ABSTRACT: Cu,Zn superoxide dismutase (Cu,ZnSOD) from the thermophilic fungus Chaetomium thermophilum was expressed in Pichia pastoris and purified. Crystals were grown in over 120 conditions but only those produced with 1.4 M sodium potassium phosphate pH 8.2 as precipitant were suitable for structural studies. Data were collected to 1.9 A resolution at 100 K from a single crystal using a synchrotron-radiation source. The crystals belonged to space group P6(1)/P6(5), with unit-cell parameters a=90.2, c=314.5 A and eight molecules in the asymmetric unit. Elucidation of the crystal structure will provide insights into the active site of the enzyme and a better understanding of the structure-activity relationship, assembly and thermal stability of Cu,ZnSODs.

SUBMITTER: Wakadkar S

PROVIDER: S-EPMC2935235 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature

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Expression, purification and crystallization of Chaetomium thermophilum Cu,Zn superoxide dismutase.

Wakadkar Sachin S Zhang Li-Qing LQ Li Duo-Chuan DC Haikarainen Teemu T Dhavala Prathusha P Papageorgiou Anastassios C AC

Acta crystallographica. Section F, Structural biology and crystallization communications 20100828 Pt 9

Cu,Zn superoxide dismutase (Cu,ZnSOD) from the thermophilic fungus Chaetomium thermophilum was expressed in Pichia pastoris and purified. Crystals were grown in over 120 conditions but only those produced with 1.4 M sodium potassium phosphate pH 8.2 as precipitant were suitable for structural studies. Data were collected to 1.9 A resolution at 100 K from a single crystal using a synchrotron-radiation source. The crystals belonged to space group P6(1)/P6(5), with unit-cell parameters a=90.2, c=31 ...[more]

PMID: 20823534

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