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Journal of Virology, November 2006, p. 10615-10623, Vol. 80, No. 21
0022-538X/06/$08.00+0     doi:10.1128/JVI.01452-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Proteomic Analysis of the Major Envelope and Nucleocapsid Proteins of White Spot Syndrome Virus{triangledown}

Xixian Xie,1,2 Limei Xu,1 and Feng Yang1*

Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, Xiamen 361005, People's Republic of China,1 School of Life Sciences, Xiamen University, Xiamen 361005, People's Republic of China2

Received 10 July 2006/ Accepted 16 August 2006

White spot syndrome virus (WSSV) virions were purified from the tissues of infected Procambarus clarkii (crayfish) isolates. Pure WSSV preparations were subjected to Triton X-100 treatment to separate into the envelope and nucleocapsid fractions, which were subsequently separated by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The major envelope and nucleocapsid proteins were identified by either matrix-assisted laser desorption ionization-time of flight mass spectrometry or defined antibody. A total of 30 structural proteins of WSSV were identified in this study; 22 of these were detected in the envelope fraction, 7 in the nucleocapsid fraction, and 1 in both the envelope and the nucleocapsid fractions. With the aid of specific antibodies, the localizations of eight proteins were further studied. The analysis of posttranslational modifications revealed that none of the WSSV structural proteins was glycosylated and that VP28 and VP19 were threonine phosphorylated. In addition, far-Western and coimmunoprecipitation experiments showed that VP28 interacted with both VP26 and VP24. In summary, the data obtained in this study should provide an important reference for future molecular studies of WSSV morphogenesis.

* Corresponding author. Mailing address: Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, SOA, 178 Daxue Rd., Xiamen 361005, People's Republic of China. Phone: 86-0592-2195274. Fax: 86-0592-2085376. E-mail: mbiotech{at}public.xm.fj.cn.

{triangledown} Published ahead of print on 23 August 2006.

Journal of Virology, November 2006, p. 10615-10623, Vol. 80, No. 21
0022-538X/06/$08.00+0     doi:10.1128/JVI.01452-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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