BIBLIOS

Document type
Journal articles

Document subtype
Full paper

Title
A Proteomic Approach toward the Selection of Proteins with Enhanced Intrinsic Conformational Stability

Participants in the publication
Vesna Prosinecki (Author)
Hugo M. Botelho (Author)
Simona Francese (Author)
Guido Mastrobuoni (Author)
Gloriano Moneti (Author)
Tim Urich (Author)
Arnulf Kletzin (Author)
Cláudio M. Gomes (Author)
Dep. Química e Bioquímica
BioISI

Summary
A detailed understanding of the molecular basis of protein folding and stability determinants partly relies on the study of proteins with enhanced conformational stability properties, such as those from thermophilic organisms. In this study, we set up a methodology aiming at identifying the subset of cytosolic hyperstable proteins using Sulfurispharea sp., a hyperthermophilic archaeon, able to grow between 70 and 97 °C, as a model organism. We have thermally and chemically perturbed the cytosolic proteome as a function of time (up to 96 h incubation at 90 °C), and proceeded with analysis of the remaining proteins by combining one- and two-dimensional gel electrophoresis, liquid chromatography fractionation, and protein identification by N-terminal sequencing and mass spectrometry methods. In total, 14 proteins with enhanced stabilities which are involved in key cellular processes such as detoxification, nucleic acid processing, and energy metabolism were identified including a superoxide dismutase, a peroxiredoxin, and a ferredoxin. We demonstrate that these proteins are biologically active after extensive thermal treatment of the proteome. The relevance of these and other targets is discussed in terms of the organism's ecology. This work thus illustrates an experimental approach aimed at mining a proteome for hyperstable proteins, a valuable tool for target selection in protein stability and structural studies.


Institution
INSTITUTO DE TECNOLOGIA QUÍMICA E BIOLÓGICA

Where published
Journal of Proteome Research

Publication Identifiers
ISSN - 1535-3893
eISSN - 1535-3907

Publisher
American Chemical Society (ACS)



Document Identifiers
DOI - https://doi.org/10.1021/pr0602491
URL - http://dx.doi.org/10.1021/pr0602491