1887

Abstract

Previous work has shown that the avian reovirus cell-attachment sigma C (σC) protein is a multimer. In the first part of this study the oligomerization state of intracellularly synthesized σC was analysed by different approaches, including SDS–PAGE, chemical cross-linking, sedimentation and gel filtration analysis. All these approaches indicated that protein σC in its native state is a homotrimer. In the second part of the present work we investigated the effect of different factors and reagents on oligomer stability, in order to elucidate the nature of the forces that maintain the conformational stability of the homotrimer. Our results, based on the stabilizing effect conferred by reducing agents, demonstrate that the σC subunits are not covalently bound via disulfide linkages. They further suggest that the formation of an intrachain disulfide bond between the two cysteine residues of the σC polypeptide has a negative effect on oligomer stability. The susceptibility of the trimer to pH, temperature, ionic strength, chemical denaturants and detergents indicates that hydrophobic interactions contribute much more to oligomer stability than do ionic interactions and hydrogen bonding. Finally, our results also reveal that mammalian and avian reovirus cell attachment proteins follow different subunit dissociation pathways.

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2002-01-01
2020-01-23
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