Thermal stability and aggregation of Sulfolobus solfataricus β-glycosidase are dependent upon the N-ε-methylation of specific lysyl residues: Critical role of in vivo post-translational modifications

Ferdinando Febbraio, Annapaola Andolfo, Fabio Tanfani, Raffaella Briante, Fabrizio Gentile, Silvestro Formisano, Carlo Vaccaro, Andrea Scirè, Enrico Bertoli, Piero Pucci, Roberto Nucci

Research output: Contribution to journalArticlepeer-review

Abstract

Methylation in vivo is a post-translational modification observed in several organisms belonging to eucarya, bacteria, and archaea. Although important implications of this modification have been demonstrated in several eucaryotes, its biological role in hyperthermophilic archaea is far from being understood. The aim of this work is to clarify some effects of methylation on the properties of β-glycosidase from Sulfolobus solfataricus, by a structural comparison between the native, methylated protein and its unmethylated counterpart, recombinantly expressed in Escherichia coli. Analysis by Fourier transform infrared spectroscopy indicated similar secondary structure contents for the two forms of the protein. However, the study of temperature perturbation by Fourier transform infrared spectroscopy and turbidimetry evidenced denaturation and aggregation events more pronounced in recombinant than in native β-glycosidase. Red Nile fluorescence analysis revealed significant differences of surface hydrophobicity between the two forms of the protein. Unlike the native enzyme, which dissociated into SDS-resistant dimers upon exposure to the detergent, the recombinant enzyme partially dissociated into monomers. By electrospray mapping, the methylation sites of the native protein were identified. A computational analysis of β-glycosidase three-dimensional structure and comparisons with other proteins from S. solfataricus revealed analogies in the localization of methylation sites in terms of secondary structural elements and overall topology. These observations suggest a role for the methylation of lysyl residues, located in selected domains, in the thermal stabilization of β-glycosidase from S. solfataricus.

Original languageEnglish
Pages (from-to)10185-10194
Number of pages10
JournalJournal of Biological Chemistry
Volume279
Issue number11
DOIs
Publication statusPublished - Mar 12 2004

ASJC Scopus subject areas

  • Biochemistry

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