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Purification, crystallization and preliminary X-ray diffraction analysis of the effector protein PevD1 from Verticillium dahliae.

ABSTRACT: The effector protein PevD1 from the pathogenic fungus Verticillium dahliae was purified and crystallized using the hanging-drop vapour-diffusion method. Native crystals appeared in a solution consisting of 4.0?M sodium formate. A native data set was collected at 1.9?Å resolution at 100?K using an in-house X-ray source. Because of the absence of useful methinione in the protein sequence, derivative crystals that contained iodine were obtained by soaking in 1.25?M potassium iodide, and a data set that contained anomalous signal was collected using the same X-ray facility at a wavelength of 1.54?Å. The single-wavelength anomalous dispersion method was used to successfully solve the structure based on the anomalous signal generated from iodine.

SUBMITTER: Han L

PROVIDER: S-EPMC3388926 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature

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Purification, crystallization and preliminary X-ray diffraction analysis of the effector protein PevD1 from Verticillium dahliae.

Han Lei L Liu Zheng Z Liu Xinqi X Qiu Dewen D

Acta crystallographica. Section F, Structural biology and crystallization communications 20120628 Pt 7

The effector protein PevD1 from the pathogenic fungus Verticillium dahliae was purified and crystallized using the hanging-drop vapour-diffusion method. Native crystals appeared in a solution consisting of 4.0 M sodium formate. A native data set was collected at 1.9 Å resolution at 100 K using an in-house X-ray source. Because of the absence of useful methinione in the protein sequence, derivative crystals that contained iodine were obtained by soaking in 1.25 M potassium iodide, and a data set ...[more]

PMID: 22750869

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