Proline 235 plays a key role in the regulation of the oligomeric states of Thermotoga maritima Arginine Binding Protein

Giovanni Smaldone, Marilisa Vigorita, Alessia Ruggiero, Nicole Balasco, Jonathan D. Dattelbaum, Sabato D'Auria, Pompea Del Vecchio, Giuseppe Graziano, Luigi Vitagliano

Research output: Contribution to journalArticle

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Abstract

The Arginine Binding Protein isolated from Thermotoga maritima (TmArgBP) is a protein endowed with several peculiar properties. We have previously shown that TmArgBP dimerization is a consequence of the swapping of the C-terminal helix. Here we explored the structural determinants of TmArgBP domain swapping and oligomerization. In particular, we report a mutational analysis of the residue Pro235, which is located in the hinge region of the swapping dimer. This residue was either replaced with a Gly-Lys dipeptide (TmArgBPP235GK) or a Gly residue (TmArgBPP235G). Different forms of these mutants were generated and extensively characterized using biophysical techniques. For both TmArgBPP235GK and TmArgBPP235G mutants, the occurrence of multiple oligomerization states (monomers, dimers and trimers) was detected. The formation of well-folded monomeric forms for these mutants indicates that the dimerization through C-terminal domain swapping observed in wild-type TmArgBP is driven by conformational restraints imposed by the presence of Pro235 in the hinge region. Molecular dynamics studies corroborate this observation by showing that Gly235 assumes conformational states forbidden for Pro residues in the TmArgBPP235G monomer. Unexpectedly, the trimeric forms present: (a) peculiar circular dichroism spectra, (b) a great susceptibility to heating, and (c) the ability to bind the Thioflavin T dye. The present findings clearly demonstrate that single-point mutations have an important impact on the TmArgBP oligomerization process. In a wider context, they also indicate that proteins endowed with an intrinsic propensity to swap have an easy access to states with altered structural and, possibly, functional properties.

Original languageEnglish
Pages (from-to)814-824
Number of pages11
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1864
Issue number7
DOIs
Publication statusPublished - Jul 1 2016

Fingerprint

Thermotoga maritima
Oligomerization
Dimerization
Proline
glycyllysine
Arginine
Carrier Proteins
Hinges
Dimers
Dipeptides
Molecular Dynamics Simulation
Circular Dichroism
Monomers
Point Mutation
Heating
Proteins
Coloring Agents
Dichroism
Molecular dynamics

Keywords

  • Differential scanning calorimetry
  • Domain swapping
  • Protein oligomerization
  • Protein structure dynamics
  • Protein structure-stability

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Proline 235 plays a key role in the regulation of the oligomeric states of Thermotoga maritima Arginine Binding Protein. / Smaldone, Giovanni; Vigorita, Marilisa; Ruggiero, Alessia; Balasco, Nicole; Dattelbaum, Jonathan D.; D'Auria, Sabato; Del Vecchio, Pompea; Graziano, Giuseppe; Vitagliano, Luigi.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1864, No. 7, 01.07.2016, p. 814-824.

Research output: Contribution to journalArticle

Smaldone, G, Vigorita, M, Ruggiero, A, Balasco, N, Dattelbaum, JD, D'Auria, S, Del Vecchio, P, Graziano, G & Vitagliano, L 2016, 'Proline 235 plays a key role in the regulation of the oligomeric states of Thermotoga maritima Arginine Binding Protein', Biochimica et Biophysica Acta - Proteins and Proteomics, vol. 1864, no. 7, pp. 814-824. https://doi.org/10.1016/j.bbapap.2016.04.006
Smaldone, Giovanni ; Vigorita, Marilisa ; Ruggiero, Alessia ; Balasco, Nicole ; Dattelbaum, Jonathan D. ; D'Auria, Sabato ; Del Vecchio, Pompea ; Graziano, Giuseppe ; Vitagliano, Luigi. / Proline 235 plays a key role in the regulation of the oligomeric states of Thermotoga maritima Arginine Binding Protein. In: Biochimica et Biophysica Acta - Proteins and Proteomics. 2016 ; Vol. 1864, No. 7. pp. 814-824.
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AU - Dattelbaum, Jonathan D.

AU - D'Auria, Sabato

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