Abstract
VP2 is the minor structural protein of noroviruses (NoV) and may function in NoV particle stability. To determine the function of VP2 in the stability of the NoV particle, we constructed and purified two kinds of virus-like particles (VLPs), namely, VLPs (VP1) and VLPs (VP1+VP2), from Sf9 cells infected with recombinant baculoviruses by using a Bac-to-Bac® baculovirus expression system. The two kinds of VLPs were treated with different phosphate buffers (pH 2 to pH 8); the secondary structure was then analyzed by far UV circular dichroism (CD) spectroscopy. Results showed that significant disruptions of the secondary structure of proteins were not observed at pH 2 to pH 7. At pH 8, the percentages of a-helix, β-sheet, and β-turn in VLPs (VP1) were decreased from 11% to 8%, from 37% to 32%, and from 20% to 16%, respectively. The percentage of coil was increased from 32% to 44%. By contrast, the percentages of α-helix, β-sheet, and β-turn in VLPs (VP1+VP2) were decreased from 11% to 10%, from 37% to 35%, and from 20% to 19%, respectively. The percentage of coil was increased from 32% to 36%. VLPs (VP1+VP2) was likely more stable than VLPs (VP1), as indicated by the percentage of the secondary structures analyzed by CD. These results suggested that VP2 could stabilize the secondary structure of VLPs under alkaline pH conditions. This study provided novel insights into the molecular mechanism of the function of VP2 in the stability of NoV particles.
Similar content being viewed by others
References
Ando T., Noel J.S., and Fankhauser R.L. 2000. Genetic classification of “Norwalk-like viruses”. J. Infect. Dis. 181, 336–348.
Atmar R.L. and Estes M.K. 2006. The epidemiologic and clinical importance of norovirus infection. Gastroenterol. Clin. N. Am. 35, 275–290.
Ausar S.F., Foubert T.R., Hudson M.H., Vedvick T.S., and Middaugh C.R. 2006. Conformational stability and disassembly of Norwalk virus-like particles: Effect of pH and temperature. J. Biol. Chem. 281, 19478–19488.
Bertolotti-Ciarlet A., Crawford S.E., Hutson A.M., and Estes M.K. 2003. The 3' end of Norwalk virus mRNA contains determinants that regulate the expression and stability of the viral capsid protein VP1: a novel function for the VP2 protein. J. Virol. 77, 11603–11605.
Bertolotti-Ciarlet A., White L.J., Chen R., Prasad B.V., and Estes M.K. 2002. Structural requirements for the assembly of Norwalk virus-like particles. J. Virol. 76, 4044–4055.
Culver J.N. 2002. Tobacco mosaic virus assembly and disassembly: determinants in pathogenicity and resistance. Annu. Rev. Phytopathol. 40, 287–308.
Doultree J.C., Druce J.D., Birch C.J., Bowden D.S., and Marshall J.A. 1999. Inactivation of feline calicivirus, a Norwalk virus surrogate. J. Hosp. Infect. 41, 51–57.
Glass P.J., White L.J., Ball J.M., Leparc-Goffart I., Hardy M.E., and Estes M.K. 2000. Norwalk virus open reading frame 3 encodes a minor structural protein. J. Virol. 74, 6581–6591.
Green K.Y., Kapikian A.Z., Valdesuso J., Sosnovtsev S., Treanor J.J., and Lew J.F. 1997. Expression and self-assembly of recombinant capsid protein from the antigenically distinct Hawaii human calicivirus. J. Clin. Microbiol. 35, 1909–1914.
Hale A.D., Crawford S.E., Ciarlet M., Green J., Gallimore C., Brown D.W., Jiang X., and Estes M.K. 1999. Expression and self-assembly of Grimsby virus: antigenic distinction from Norwalk and Mexico viruses. Clin. Diagn. Lab. Immunol. 6, 142–145.
Jiang X., Matson D.O., Ruiz-Palacios G.M., Hu J., Treanor J., and Pickering L.K. 1995. Expression, self-assembly, and antigenicity of a snow mountain agent-like calicivirus capsid protein. J. Clin. Microbiol. 33, 1452–1455.
Jiang X., Wang M., Graham D.Y., and Estes M.K. 1992. Expression, self-assembly, and antigenicity of the Norwalk virus capsid protein. J. Virol. 66, 6527–6532.
Jiang X., Wang M., Wang K., and Estes M.K. 1993. Sequence and genomic organization of Norwalk virus. Virology 195, 51–61.
Karst S.M., Wobus C.E., Lay M., Davidson J., and Virgin H.W.IV. 2003. STAT1-dependent innate immunity to a Norwalk-like virus. Science 299, 1575–1578.
Kobayashi S., Sakae K., Suzuki Y., Shinozaki K., Okada M., Ishiko H., Kamata K., Suzuki K., Natori K., Miyamura T., and Takeda N. 2000. Molecular cloning, expression, and antigenicity of Seto virus belonging to genogroup I Norwalk-like viruses. J. Clin. Microbiol. 38, 3492–3494.
Kroneman A., Harris J.R., Vennema H., Duizer E., van Duynhoven Y., Gray J., Iturriza M., Bottiger B., Falkenhorst G., Johnsen C., and et al. 2008. Data quality of 5 years of central norovirus outbreak reporting in the European Network for food-borne viruses. J. Public Health 30, 82–90.
Lindesmith L., Moe C., Marionneau S., Ruvoen N., Jiang X., Lindblad L., Stewart P., LePendu J., and Baric R. 2003. Human susceptibility and resistance to Norwalk virus infection. Nat. Med. 9, 548–553.
Luttermann C. and Meyers G. 2007. A bipartite sequence motif induces translation reinitiation in feline calicivirus RNA. J. Biol. Chem. 282, 7056–7065.
Marionneau S., Ruvoën N., Le Moullac-Vaidye B., Clement M., Cailleau-Thomas A., Ruiz-Palacois G., Huang P., Jiang X., and Le Pendu J. 2002. Norwalk virus binds to histo-blood group antigens present on gastroduodenal epithelial cells of secretor individuals. Gastroenterology 122, 1967–1977.
Pirtle E.C. and Beran G.W. 1991. Virus survival in the environment. Rev. Sci. Tech. 10, 733–748.
Prasad B.V., Hardy M.E., Dokland T., Bella J., Rossmann M.G., and Estes M.K. 1999. X-ray crystallographic structure of the Norwalk virus capsid. Science 286, 287–290.
Sosnovtsev S.V., Belliot G., Chang K.O., Onwudiwe O., and Green K.Y. 2005. Feline calicivirus VP2 is essential for the production of infectious virions. J. Virol. 79, 4012–4024.
Svraka S., Duizer E., Vennema H., de Bruin E., van der Veer B., Dorresteijn B., and Koopmans M. 2007. Etiological role of viruses in outbreaks of acute gastroenteritis in The Netherlands from 1994 through 2005. J. Clin. Microbiol. 45, 1389–1394.
Vongpunsawad S., Venkataram Prasad B.V., and Estes M.K. 2013. Norwalk virus minor capsid protein VP2 associates within the VP1 shell domain. J. Virol. 87, 4818–4825.
Wang D., Wu Q., Kou X., Yao L., and Zhang J. 2008. Distribution of norovirus in oyster tissues. J. Appl. Microbiol. 105, 1966–1972.
White L.J., Hardy M.E., and Estes M.K. 1997. Biochemical characterization of a smaller form of recombinant Norwalk virus capsids assembled in insect cells. J. Virol. 71, 8066–8072.
Widdowson M.A., Monroe S.S., and Glass R.I. 2005. Are noroviruses emerging? Emerg. Infect. Dis. 11, 735–737.
Yang J.T., Wu C.S.C., and Martinez H.M. 1986. Calculation of protein conformation from circular dichroism. Methods Enzymol. 130, 208–269.
Zheng D.P., Ando T., Fankhauser R.L., Beard R.S., Glass R.I., and Monroe S.S. 2006. Norovirus classification and proposed strain nomenclature. Virology 346, 312–323.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lin, Y., Fengling, L., Lianzhu, W. et al. Function of VP2 protein in the stability of the secondary structure of virus-like particles of genogroup II norovirus at different pH levels: Function of VP2 protein in the stability of NoV VLPs. J Microbiol. 52, 970–975 (2014). https://doi.org/10.1007/s12275-014-4323-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12275-014-4323-6