Thermal stability and aggregation of Sulfolobus solfataricus beta-glycosidase are dependent upon N-epsilon-methylation of specific lysyl residues

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 to be understood. The aim of this work is to clarify some effects of methylation on the properties of beta-glycosidase from Sulfolobus solfataricus, by a structural comparison between the native, methylated protein and its unmethylated counterpart, recombinantly expressed in E. coli. Analysis by FT-IR spectroscopy indicated similar secondary structure contents for the two forms of the protein. However, the study of temperature perturbation by FT-IR spectroscopy and turbidimetry evidenced denaturation and aggregation events more pronounced in recombinant than in native beta-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 beta-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 beta-glycosidase from S. solfataricus.