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Expression, purification and crystallization of MnSOD from Arabidopsis thaliana
Author(s) -
Marques Alexandra T.,
Santos Sandra P.,
Rosa Margarida G.,
Rodrigues Mafalda A. A.,
Abreu Isabel A.,
Frazão Carlos,
Romão Célia V.
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14007687
Subject(s) - arabidopsis thaliana , molecular replacement , arabidopsis , crystallization , abiotic stress , chemistry , biology , crystallography , biochemistry , enzyme , gene , organic chemistry , mutant
Manganese superoxide dismutase (MnSOD) is an essential primary antioxidant enzyme. MnSOD plays an important role in plant tolerance to abiotic stress and is a target candidate for increasing stress tolerance in crop plants. Although the structure and kinetic parameters of MnSODs from several organisms have been determined, this information is still lacking for plant MnSODs. Here, recombinant MnSOD from Arabidopsis thaliana (AtMnSOD) was expressed, purified and crystallized. A nearly complete data set could only be obtained when a total rotation range of 180° was imposed during data collection, despite the seemingly tetragonal metric of the AtMnSOD crystal diffraction. The data set extended to 1.95 Å resolution and the crystal belonged to space group P 1. Molecular‐replacement calculations using an ensemble of homologous SOD structures as a search model gave a unique and unambiguous solution corresponding to eight molecules in the asymmetric unit. Structural and kinetic analysis of AtMnSOD is currently being undertaken.