NMR studies on structure and dynamics of the monomeric derivative of BS-RNase: New insights for 3D domain swapping

Roberta Spadaccini, Carmine Ercole, Maria A. Gentile, Domenico Sanfelice, Rolf Boelens, Rainer Wechselberger, G. Batta, Andrea Bernini, Neri Niccolai, Delia Picone

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Three-dimensional domain swapping is a common phenomenon in pancreatic-like ribonucleases. In the aggregated state, these proteins acquire new biological functions, including selective cytotoxicity against tumour cells. RNase A is able to dislocate both N- and C-termini, but usually this process requires denaturing conditions. In contrast, bovine seminal ribonuclease (BS-RNase), which is a homo-dimeric protein sharing 80% of sequence identity with RNase A, occurs natively as a mixture of swapped and unswapped isoforms. The presence of two disulfides bridging the subunits, indeed, ensures a dimeric structure also to the unswapped molecule. In vitro, the two BS-RNase isoforms interconvert under physiological conditions. Since the tendency to swap is often related to the instability of the monomeric proteins, in these paper we have analysed in detail the stability in solution of the monomeric derivative of BS-RNase (mBS) by a combination of NMR studies and Molecular Dynamics Simulations. The refinement of NMR structure and relaxation data indicate a close similarity with RNase A, without any evidence of aggregation or partial opening. The high compactness of mBS structure is confirmed also by H/D exchange, urea denaturation, and TEMPOL mapping of the protein surface. The present extensive structural and dynamic investigation of (monomeric) mBS did not show any experimental evidence that could explain the known differences in swapping between BS-RNase and RNase A. Hence, we conclude that the swapping in BS-RNase must be influenced by the distinct features of the dimers, suggesting a prominent role for the interchain disulfide bridges.

Original languageEnglish
Article numbere29076
JournalPLoS One
Volume7
Issue number1
DOIs
Publication statusPublished - Jan 12 2012

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Pancreatic Ribonuclease
ribonucleases
chemical derivatives
Nuclear magnetic resonance
Derivatives
Ribonucleases
cattle
Disulfides
sulfides
Protein Isoforms
proteins
Proteins
Denaturation
Cytotoxicity
molecular dynamics
Dimers
Homo
Molecular Dynamics Simulation
Urea
Molecular dynamics

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Spadaccini, R., Ercole, C., Gentile, M. A., Sanfelice, D., Boelens, R., Wechselberger, R., ... Picone, D. (2012). NMR studies on structure and dynamics of the monomeric derivative of BS-RNase: New insights for 3D domain swapping. PLoS One, 7(1), [e29076]. https://doi.org/10.1371/journal.pone.0029076

NMR studies on structure and dynamics of the monomeric derivative of BS-RNase : New insights for 3D domain swapping. / Spadaccini, Roberta; Ercole, Carmine; Gentile, Maria A.; Sanfelice, Domenico; Boelens, Rolf; Wechselberger, Rainer; Batta, G.; Bernini, Andrea; Niccolai, Neri; Picone, Delia.

In: PLoS One, Vol. 7, No. 1, e29076, 12.01.2012.

Research output: Contribution to journalArticle

Spadaccini, R, Ercole, C, Gentile, MA, Sanfelice, D, Boelens, R, Wechselberger, R, Batta, G, Bernini, A, Niccolai, N & Picone, D 2012, 'NMR studies on structure and dynamics of the monomeric derivative of BS-RNase: New insights for 3D domain swapping', PLoS One, vol. 7, no. 1, e29076. https://doi.org/10.1371/journal.pone.0029076
Spadaccini R, Ercole C, Gentile MA, Sanfelice D, Boelens R, Wechselberger R et al. NMR studies on structure and dynamics of the monomeric derivative of BS-RNase: New insights for 3D domain swapping. PLoS One. 2012 Jan 12;7(1). e29076. https://doi.org/10.1371/journal.pone.0029076
Spadaccini, Roberta ; Ercole, Carmine ; Gentile, Maria A. ; Sanfelice, Domenico ; Boelens, Rolf ; Wechselberger, Rainer ; Batta, G. ; Bernini, Andrea ; Niccolai, Neri ; Picone, Delia. / NMR studies on structure and dynamics of the monomeric derivative of BS-RNase : New insights for 3D domain swapping. In: PLoS One. 2012 ; Vol. 7, No. 1.
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