Two Novel Amyloid Proteins, RopA and RopB, from the Root Nodule Bacterium Rhizobium leguminosarum

Anastasiia O. Kosolapova, Mikhail V. Belousov, Anna I. Sulatskaya, Maria E. Belousova, Maksim I. Sulatsky, Kirill S. Antonets, Kirill V. Volkov, Anna N. Lykholay, Oksana Y. Shtark, Ekaterina N. Vasileva, Vladimir A. Zhukov, Alexandra N. Ivanova, Pavel A. Zykin, Irina M. Kuznetsova, Konstantin K. Turoverov, Igor A. Tikhonovich, Anton A. Nizhnikov

Research output

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Abstract

Amyloids represent protein fibrils with a highly ordered spatial structure, which not only cause dozens of incurable human and animal diseases but also play vital biological roles in Archaea, Bacteria, and Eukarya. Despite the fact that association of bacterial amyloids with microbial pathogenesis and infectious diseases is well known, there is a lack of information concerning the amyloids of symbiotic bacteria. In this study, using the previously developed proteomic method for screening and identification of amyloids (PSIA), we identified amyloidogenic proteins in the proteome of the root nodule bacterium Rhizobium leguminosarum. Among 54 proteins identified, we selected two proteins, RopA and RopB, which are predicted to have β-barrel structure and are likely to be involved in the control of plant-microbial symbiosis. We demonstrated that the full-length RopA and RopB form bona fide amyloid fibrils in vitro. In particular, these fibrils are β-sheet-rich, bind Thioflavin T (ThT), exhibit green birefringence upon staining with Congo Red (CR), and resist treatment with ionic detergents and proteases. The heterologously expressed RopA and RopB intracellularly aggregate in yeast and assemble into amyloid fibrils at the surface of Escherichia coli. The capsules of the R. leguminosarum cells bind CR, exhibit green birefringence, and contain fibrils of RopA and RopB in vivo.

Original languageEnglish
Article number694
Number of pages25
JournalBiomolecules
Volume9
Issue number11
DOIs
Publication statusPublished - 4 Nov 2019

Scopus subject areas

  • Biochemistry
  • Molecular Biology

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  • Cite this

    Kosolapova, A. O., Belousov, M. V., Sulatskaya, A. I., Belousova, M. E., Sulatsky, M. I., Antonets, K. S., Volkov, K. V., Lykholay, A. N., Shtark, O. Y., Vasileva, E. N., Zhukov, V. A., Ivanova, A. N., Zykin, P. A., Kuznetsova, I. M., Turoverov, K. K., Tikhonovich, I. A., & Nizhnikov, A. A. (2019). Two Novel Amyloid Proteins, RopA and RopB, from the Root Nodule Bacterium Rhizobium leguminosarum. Biomolecules, 9(11), [694]. https://doi.org/10.3390/biom9110694