Abstract
The advent of native mass spectrometry (MS) in 1990 led to the development of new mass spectrometry instrumentation and methodologies for the analysis of noncovalent protein–ligand complexes. Native MS has matured to become a fast, simple, highly sensitive and automatable technique with well-established utility for fragment-based drug discovery (FBDD). Native MS has the capability to directly detect weak ligand binding to proteins, to determine stoichiometry, relative or absolute binding affinities and specificities. Native MS can be used to delineate ligand-binding sites, to elucidate mechanisms of cooperativity and to study the thermodynamics of binding. This review highlights key attributes of native MS for FBDD campaigns.
Highlights
In 1989 the scientific community witnessed what was thought to be impossible: “flying elephants”, as was later described by John Fenn in his Nobel Lecture [1]
They showed that electrosprayed cytochrome c molecules assumed wider distributions of charge states centered around higher charge states when sprayed from pH 2.6 than from pH 5.2 aqueous solutions, arguing this resulted from differences in the conformational states of the protein in solution
The results show that Plasmodium vivax guanylate kinase (PvGK) assumes a near-native conformation in the gas-phase, where in the absence of water and hydrophobic interactions, electrostatic interactions drive the structure to close down even without substrate binding
Summary
In 1989 the scientific community witnessed what was thought to be impossible: “flying elephants”, as was later described by John Fenn in his Nobel Lecture [1]. The first example of the application of the direct ESI-MS assay to quantify protein-ligand characterized noncovalent complexes of various modified benzensulphonamide inhibitors and affinities was reported in 1993 by Loo and co-workers [15] Another important contribution carbonic anhydrase (CA) by ESI-FT-ICR (Fourier Transform-Ion Cyclotron Resonance) [16]. A versatile instrument that geometry were developed, exploiting the m/z-range benefits of time-of-flight with the selection has great mass accuracy and resolution Another boost for the analysis of macromolecular protein abilities of a quadrupole [8]. Fourier transform ion cyclotron resonance (FT-ICR) is a versatile instrument that has great mass accuracy and resolution Another boost for the analysis of macromolecular protein complexes was the introduction of nano-ESI. Concerns commonly raised about this technique and challenges inherent to its application in FBDD are discussed
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