Structural bases for the regulation of CO binding in the archaeal protoglobin from Methanosarcina acetivorans

Lesley Tilleman, Stefania Abbruzzetti, Chiara Ciaccio, Giampiero De Sanctis, Marco Nardini, Alessandra Pesce, Filip Desmet, Luc Moens, Sabine Va Doorslaer, Stefano Bruno, Martino Bolognesi, Paolo Ascenzi, Massimo Coletta, Cristiano Viappiani, Sylvia Dewilde

Research output: Contribution to journalArticlepeer-review

Abstract

Studies of CO ligand binding revealed that two protein states with different ligand affinities exist in the protoglobin from Methanosarcina acetivorans (in MaPgb∗, residue Cys(E20)101 was mutated to Ser). The switch between the two states occurs upon the ligation of MaPgb∗. In this work, site-directed mutagenesis was used to explore the role of selected amino acids in ligand sensing and stabilization and in affecting the equilibrium between the "more reactive" and "less reactive" conformational states of MaPgb∗. A combination of experimental data obtained from electronic and resonance Raman absorption spectra, CO ligand-binding kinetics, and X-ray crystallography was employed. Three amino acids were assigned a critical role: Trp(60)B9, Tyr(61)B10, and Phe(93)E11. Trp(60)B9 and Tyr(61) B10 are involved in ligand stabilization in the distal heme pocket; the strength of their interaction was reflected by the spectra of the CO-ligated MaPgb∗and by the CO dissociation rate constants. In contrast, Phe(93)E11 is a key player in sensing the heme-bound ligand and promotes the rotation of the Trp(60)B9 side chain, thus favoring ligand stabilization. Although the structural bases of the fast CO binding rate constant of MaPgb∗are still unclear, Trp(60)B9, Tyr(61)B10, and Phe(93)E11 play a role in regulating heme/ligand affinity.

Original languageEnglish
Article numbere0125959
JournalPLoS One
Volume10
Issue number6
DOIs
Publication statusPublished - Jun 5 2015

ASJC Scopus subject areas

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

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