Abstract
Salicylic acid (SA) is an important phytohormone that is involved in the regulation of plant defence, growth and development. A large number of proteins have been shown to have the ability to interact with SA, and NPR4 has been demonstrated to be a receptor of SA that plays significant roles in the innate immune response of plants. In this study, Spodoptera frugiperda (Sf9) cells were used to express full-length AtNPR4 from Arabidopsis thaliana. To facilitate crystallization, T4 lysozyme (T4L) was added to the N-terminus of the AtNPR4 protein. The recombinant T4L-AtNPR4 protein was expressed, purified and crystallized using the sitting-drop and hanging-drop vapour-diffusion methods. The T4L-AtNPR4 crystals have symmetry consistent with space group C2, with unit-cell parameters a = 93.7, b = 85.8, c = 88.2 Å, β = 90° and one molecule per asymmetric unit. The best crystal diffracted to a resolution of 2.75 Å. Structure determination is in progress.
Keywords:
Arabidopsis thaliana; crystal diffraction; protein purification; regulatory mechanism; salicylic acid; systemic acquired resistance.
MeSH terms
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Amino Acid Sequence
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Animals
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Arabidopsis / chemistry*
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Arabidopsis / genetics
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Arabidopsis / immunology
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Arabidopsis Proteins / chemistry*
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Arabidopsis Proteins / genetics
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Arabidopsis Proteins / immunology
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Bacteriophage T4 / chemistry
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Bacteriophage T4 / genetics
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Bacteriophage T4 / metabolism
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Baculoviridae / genetics
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Baculoviridae / metabolism
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Cloning, Molecular
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Crystallization
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Crystallography, X-Ray
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Gene Expression
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Genetic Vectors / chemistry
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Genetic Vectors / metabolism
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Muramidase / chemistry
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Muramidase / genetics
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Muramidase / metabolism
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Plant Immunity / genetics
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Protein Isoforms / chemistry
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Protein Isoforms / genetics
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Protein Isoforms / immunology
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Recombinant Fusion Proteins / chemistry*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Salicylic Acid / chemistry
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Salicylic Acid / immunology
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Salicylic Acid / metabolism
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Sequence Alignment
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Sequence Homology, Amino Acid
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Sf9 Cells
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Spodoptera
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Viral Proteins / chemistry
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Viral Proteins / genetics
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Viral Proteins / metabolism
Substances
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Arabidopsis Proteins
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NPR4 protein, Arabidopsis
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Protein Isoforms
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Recombinant Fusion Proteins
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Recombinant Proteins
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Viral Proteins
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Muramidase
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Salicylic Acid