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

Anastasiia O. Kosolapova , Mikhail V. Belousov, Anna I. Sulatskaya 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

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
DOIs
Publication statusPublished - 4 Nov 2019

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Rhizobium leguminosarum
Amyloidogenic Proteins
Amyloid
Bacteria
Birefringence
Congo Red
Animal Diseases
Symbiosis
Archaea
Proteome
Eukaryota
Detergents
Proteomics
Yeast
Escherichia coli
Capsules
Communicable Diseases
Screening
Animals
Proteins

Cite this

Kosolapova , Anastasiia O. ; Belousov, Mikhail V. ; Sulatskaya, Anna I. Sulatskaya ; Belousova, Maria E. ; Sulatsky, Maksim I. ; Antonets, Kirill S. ; Volkov, Kirill V. ; Lykholay, Anna N. ; Shtark, Oksana Y. ; Vasileva, Ekaterina N. ; Zhukov , Vladimir A. ; Ivanova, Alexandra N. ; Zykin, Pavel A. ; Kuznetsova , Irina M. ; Turoverov, Konstantin K. ; Tikhonovich, Igor A. ; Nizhnikov, Anton A. . / Two Novel Amyloid Proteins, RopA and RopB, from the Root Nodule Bacterium Rhizobium leguminosarum. In: Biomolecules. 2019 ; Vol. 9.
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title = "Two Novel Amyloid Proteins, RopA and RopB, from the Root Nodule Bacterium Rhizobium leguminosarum",
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.",
keywords = "Amyloid, Fibril, RopA, RopB, outer membrane protein, Porin, root nodule, Bacteria, plant–microbial interaction, Rhizobium leguminosarum",
author = "Kosolapova, {Anastasiia O.} and Belousov, {Mikhail V.} and Sulatskaya, {Anna I. Sulatskaya} and Belousova, {Maria E.} and Sulatsky, {Maksim I.} and Antonets, {Kirill S.} and Volkov, {Kirill V.} and Lykholay, {Anna N.} and Shtark, {Oksana Y.} and Vasileva, {Ekaterina N.} and Zhukov, {Vladimir A.} and Ivanova, {Alexandra N.} and Zykin, {Pavel A.} and Kuznetsova, {Irina M.} and Turoverov, {Konstantin K.} and Tikhonovich, {Igor A.} and Nizhnikov, {Anton A.}",
year = "2019",
month = "11",
day = "4",
doi = "10.3390/biom9110694",
language = "English",
volume = "9",
journal = "Biomolecules",
issn = "2218-273X",
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Kosolapova , AO, Belousov, MV, Sulatskaya, AIS, Belousova, ME, Sulatsky, MI, Antonets, KS, Volkov, KV, Lykholay, AN, Shtark, OY, Vasileva, EN, Zhukov , VA, Ivanova, AN, Zykin, PA, Kuznetsova , IM, Turoverov, KK, Tikhonovich, IA & Nizhnikov, AA 2019, 'Two Novel Amyloid Proteins, RopA and RopB, from the Root Nodule Bacterium Rhizobium leguminosarum', Biomolecules, vol. 9, 694. https://doi.org/10.3390/biom9110694

Two Novel Amyloid Proteins, RopA and RopB, from the Root Nodule Bacterium Rhizobium leguminosarum. / Kosolapova , Anastasiia O.; Belousov, Mikhail V.; Sulatskaya, Anna I. Sulatskaya; Belousova, Maria E. ; Sulatsky, Maksim I. ; Antonets, Kirill S. ; Volkov, Kirill V. ; Lykholay, Anna N. ; Shtark, Oksana Y. ; Vasileva, Ekaterina N. ; Zhukov , Vladimir A. ; Ivanova, Alexandra N. ; Zykin, Pavel A. ; Kuznetsova , Irina M. ; Turoverov, Konstantin K. ; Tikhonovich, Igor A. ; Nizhnikov, Anton A. .

In: Biomolecules, Vol. 9, 694, 04.11.2019.

Research output

TY - JOUR

T1 - Two Novel Amyloid Proteins, RopA and RopB, from the Root Nodule Bacterium Rhizobium leguminosarum

AU - Kosolapova , Anastasiia O.

AU - Belousov, Mikhail V.

AU - Sulatskaya, Anna I. Sulatskaya

AU - Belousova, Maria E.

AU - Sulatsky, Maksim I.

AU - Antonets, Kirill S.

AU - Volkov, Kirill V.

AU - Lykholay, Anna N.

AU - Shtark, Oksana Y.

AU - Vasileva, Ekaterina N.

AU - Zhukov , Vladimir A.

AU - Ivanova, Alexandra N.

AU - Zykin, Pavel A.

AU - Kuznetsova , Irina M.

AU - Turoverov, Konstantin K.

AU - Tikhonovich, Igor A.

AU - Nizhnikov, Anton A.

PY - 2019/11/4

Y1 - 2019/11/4

N2 - 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.

AB - 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.

KW - Amyloid

KW - Fibril

KW - RopA

KW - RopB

KW - outer membrane protein

KW - Porin

KW - root nodule

KW - Bacteria

KW - plant–microbial interaction

KW - Rhizobium leguminosarum

U2 - 10.3390/biom9110694

DO - 10.3390/biom9110694

M3 - Article

VL - 9

JO - Biomolecules

JF - Biomolecules

SN - 2218-273X

M1 - 694

ER -