Functional and structural roles of the N-terminal extension in Methanosarcina acetivorans protoglobin

Chiara Ciaccio, Alessandra Pesce, Grazia R. Tundo, Lesley Tilleman, Laura Bertolacci, Sylvia Dewilde, Luc Moens, Paolo Ascenzi, Martino Bolognesi, Marco Nardini, Massimo Coletta

Research output: Contribution to journalArticlepeer-review

Abstract

Functional and structural properties of protoglobin from Methanosarcina acetivorans, whose Cys(101)E20 residue was mutated to Ser (MaPgb *), and of mutants missing either the first 20 N-terminal amino acids (MaPgb*-ΔN20 mutant), or the first 33 N-terminal amino acids [N-terminal loop of 20 amino acids and a 13-residue Z-helix, preceding the globin fold A-helix; (MaPgb*-ΔN20Z mutant)] have been investigated. In keeping with the MaPgb*-ΔN20 mutant crystal structure, here reported at 2.0 Å resolution, which shows an increased exposure of the haem propionates to the solvent, the analysis of ligand binding kinetics highlights high accessibility ofligands tothe haem pocket inferricMaPgb*-ΔN20. CObinding to ferrous MaPgb*-ΔN20 displays a marked biphasic behavior, with a fast binding process close to that observed in MaPgb* and a slow carbonylation process, characterized by a rate-limiting step. Conversely, removal of the first 33 residues induces a substantial perturbation of the overall MaPgb* structure, with loss of α-helical content and potential partial collapse of the protein chain. As such, ligand binding kinetics are characterized by very slow rates that are independent of ligand concentration, this being indicative of a high energy barrier for ligand access to the haem, possibly due to localized misfolding. This article is part of a Special Issue entitled: Oxygen Binding and Sensing Proteins.

Original languageEnglish
Pages (from-to)1813-1823
Number of pages11
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1834
Issue number9
DOIs
Publication statusPublished - 2013

Keywords

  • Ligand binding property
  • Methanosarcina acetivorans protoglobin
  • N-terminal trimming
  • Spectroscopic property
  • Three-dimensional structure

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Analytical Chemistry
  • Molecular Biology
  • Medicine(all)

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