Conformational flexibility of proteins involved in ribosome biogenesis: Investigations via small angle x-ray scattering (SAXS)

Dritan Siliqi, James Foadi, Marco Mazzorana, Davide Altamura, Alfonso Méndez-Godoy, Nuria Sánchez-Puig

Research output: Contribution to journalArticlepeer-review

4 Citations (SciVal)

Abstract

The dynamism of proteins is central to their function, and several proteins have been described as flexible, as consisting of multiple domains joined by flexible linkers, and even as intrinsically disordered. Several techniques exist to study protein structures, but small angle X-ray scattering (SAXS) has proven to be particularly powerful for the quantitative analysis of such flexible systems. In the present report, we have used SAXS in combination with X-ray crystallography to highlight their usefulness at characterizing flexible proteins, using as examples two proteins involved in different steps of ribosome biogenesis. The yeast BRCA2 and CDKN1A-interactig protein, Bcp1, is a chaperone for Rpl23 of unknown structure. We showed that it consists of a rigid, slightly elongated protein, with a secondary structure comprising a mixture of alpha helices and beta sheets. As an example of a flexible molecule, we studied the SBDS (Shwachman-Bodian-Diamond Syndrome) protein that is involved in the cytoplasmic maturation of the 60S subunit and constitutes the mutated target in the Shwachman-Diamond Syndrome. In solution, this protein coexists in an ensemble of three main conformations, with the N- and C-terminal ends adopting different orientations with respect to the central domain. The structure observed in the protein crystal corresponds to an average of those predicted by the SAXS flexibility analysis.

Original languageEnglish
Article number109
JournalCrystals
Volume8
Issue number3
DOIs
Publication statusPublished - 1 Mar 2018

Funding

N.S.-P acknowledges the financial support from CONACyT project number 283909 and DGAPA-PAPIIT IN201615. D.S. acknowledges the financial support from projects “PGR2014-2017 Messico” with the contribution of the Ministero degli Affari Esteri e dalla Cooperazione Internazionale, Direzione Generale per la Promozione del Sistema Paese. The SAXS experiments were performed by proposal SM14029-1 and SM16970-3 beamline B21, Diamond Light Source. MX experiments were performed by proposal MX11690-13 at beamline I24, Diamond Light Source.

Keywords

  • Bcp1
  • Flexibility
  • SAXS
  • SBDS

ASJC Scopus subject areas

  • General Chemical Engineering
  • General Materials Science
  • Condensed Matter Physics
  • Inorganic Chemistry

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