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Envelope protein VP24 from White spot syndrome virus: expression, purification and crystallization.


ABSTRACT: White spot syndrome virus (WSSV) is a major shrimp pathogen known to infect penaeid shrimp and other crustaceans. VP24 is one of the major envelope proteins of WSSV. In order to facilitate purification, crystallization and structure determination, the predicted N-terminal transmembrane region of approximately 26 amino acids was truncated from VP24 and several mutants were prepared to increase the proportion of selenomethionine (SeMet) residues for subsequent structural determination using the SAD method. Truncated VP24, its mutants and the corresponding SeMet-labelled proteins were purified, and the native and SeMet proteins were crystallized by the hanging-drop vapour-diffusion method. Crystals of VP24 were obtained using a reservoir consisting of 0.1?M Tris-HCl pH 8.5, 2.75?M ammonium acetate with a drop volume ratio of two parts protein solution to one part reservoir solution. Notably, ATP was added as a critical additive to the drop with a final concentration of 10?mM. Crystals of SeMet-labelled VP24 mutant diffracted to 3.0?Å resolution and those of the native diffracted to 2.4?Å resolution; the crystals belonged to space group I213, with unit-cell parameters a = b = c = 140?Å.

SUBMITTER: Sun L 

PROVIDER: S-EPMC4973298 | BioStudies | 2016-01-01

REPOSITORIES: biostudies

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