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Comparative Analysis of Aggregation of Thermus thermophilus Ribosomal Protein bS1 and Its Stable Fragment

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Abstract

Functionally important multidomain bacterial protein bS1 is the largest ribosomal protein of subunit 30S. It interacts with both mRNA and proteins and is prone to aggregation, although this process has not been studied in detail. Here, we obtained bacterial strains overproducing ribosomal bS1 protein from Thermus thermophilus and its stable fragment bS1(49) and purified these proteins. Using fluorescence spectroscopy, dynamic light scattering, and high-performance liquid chromatography combined with mass spectrometric analysis of products of protein limited proteolysis, we demonstrated that disordered regions at the N- and C-termini of bS1 can play a key role in the aggregation of this protein. The truncated fragment bS1(49) was less prone to aggregation compared to the full-size bS1. The revealed properties of the studied proteins can be used to obtain protein crystals for elucidating the structure of the bS1 stable fragment.

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Abbreviations

bS1(49):

a fragment of Thermus thermophilus bS1 protein with a molecular mass of ∼49 kDa

CD:

circular dichroism

CSD:

cold shock domain

DLS:

dynamic light scattering

EM:

electron microscopy

HPLC/MS:

high-performance liquid chromatography/mass spectrometry

I320/I380 :

ratio of fluorescence intensities at 320 and 380 nm

m/z :

mass-to-charge ratio

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Authors and Affiliations

  1. Institute of Protein Research, Russian Academy of Sciences, 142290, Pushchino, Moscow Region, Russia

    S. Yu. Grishin, U. F. Dzhus, O. M. Selivanova, V. A. Balobanov, A. K. Surin & O. V. Galzitskaya

  2. State Research Center for Applied Microbiology and Biotechnology, 142279, Obolensk, Moscow Region, Russia

    A. K. Surin

  3. Branch of the Institute of Bioorganic Chemistry, Russian Academy of Sciences, 142290, Pushchino, Moscow Region, Russia

    A. K. Surin

  4. Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, 142290, Pushchino, Moscow Region, Russia

    O. V. Galzitskaya

Authors
  1. S. Yu. Grishin
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  2. U. F. Dzhus
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  3. O. M. Selivanova
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  4. V. A. Balobanov
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  5. A. K. Surin
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  6. O. V. Galzitskaya
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Corresponding author

Correspondence to O. V. Galzitskaya.

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Funding

This work was supported by the Russian Science Foundation (project 18-14-00321) with participation of the Center for Collective Use “Structural and Functional Studies of Proteins and RNA” of the Institute of Protein Research, Russian Academy of Sciences (584307). EM studies were carried out using equipment of the Center for Collective Use (no. 670266).

Conflict of interest

The authors declare no conflict of interest.

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This article does not contain any research involving humans or animals as research subjects performed by any of the authors.

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Grishin, S.Y., Dzhus, U.F., Selivanova, O.M. et al. Comparative Analysis of Aggregation of Thermus thermophilus Ribosomal Protein bS1 and Its Stable Fragment. Biochemistry Moscow 85, 344–354 (2020). https://doi.org/10.1134/S0006297920030104

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  • DOI: https://doi.org/10.1134/S0006297920030104

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