Electrospray‐ionization mass spectrometry as a tool for fast screening of protein structural properties

Abstract

Since the early 1990s, electrospray-ionization mass spectrometry (ESI-MS) has encountered growing interest as a complementary tool to established biochemical and biophysical methods for investigating protein structure and conformation. Nowadays, applications of ESI-MS to protein investigation span from the area of analytical biochemistry to that of structural biology. This review focuses on applications of this technique to the analysis of protein conformational properties and molecular interactions, underscoring their possible relevance for molecular biotechnology, although representing a still very young field. An introductive section presents the major issues related to theoretical and technical aspects of ESI-MS under non-denaturing conditions. Examples from our work and from the literature illustrate which kind of information can be obtained concerning key issues in biotechnology such as stability and aggregation of proteins under both near-native and challenging conditions, and interactions with other proteins, ligands and cofactors.

Structural investigation of the cold-adapted acylaminoacyl peptidase from Sporosarcina psychrophila by atomistic simulations and biophysical methods

“…Nano-ESI is a mild ionization technique that preserves non-covalent interactions and is therefore suitable for the analysis of protein conformation or supramolecular complexes [62]. The capability to preserve non-covalent interactions and the strong analytical power of mass spectrometry make nano-ESI a powerful tool for intact protein analysis [63,64]. Fig.…”

Relevance of metal ions for lipase stability: Structural rearrangements induced in the Burkholderia glumae lipase by calcium depletion

“…at high temperature. To clarify this issue we applied ESI-MS, a technique particular suited for the investigation of dynamic events occurring in metalloproteins, thanks to the stability of protein-metal complexes in the gas phase and to the possibility to monitor mass shift and conformational changes at the same time (Kaltashov et al, 2006;Grandori et al, 2009). In preliminary experiments, BGL was treated with 1% formic acid in the absence of EDTA in order to gain a reference spectrum of the denatured apo-protein.…”

Influence of acid-induced conformational variability on protein separation in reversed phase high performance liquid chromatography

“…Mass spectrometry (MS) has been widely applied for detection of species separated by HLPC 83 with on-line coupling (HPLC-MS). Besides being an analytical tool, mass spectrometry also 84 gives information on protein conformation [28][29][30][31] and on the structure of non-covalent 85 protein complexes [32,33] …”