Abstract
Analysis of intact proteins by native mass spectrometry has emerged as a powerful tool for obtaining insight into subunit diversity, post-translational modifications, stoichiometry, structural arrangement, stability, and overall architecture. Typically, such an analysis is performed following protein purification procedures, which are time consuming, costly, and labor intensive. As this technology continues to move forward, advances in sample handling and instrumentation have enabled the investigation of intact proteins in situ and in crude samples, offering rapid analysis and improved conservation of the biological context. This emerging field, which involves various ion source platforms such as matrix-assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) for both spatial imaging and solution-based analysis, is expected to impact many scientific fields, including biotechnology, pharmaceuticals, and clinical sciences. In this Perspective, we discuss the information that can be retrieved by such experiments as well as the current advantages and technical challenges associated with the different sampling strategies. Furthermore, we present future directions of these MS-based methods, including current limitations and efforts that should be made to make these approaches more accessible. Considering the vast progress we have witnessed in recent years, we anticipate that the advent of further innovations enabling minimal handling of MS samples will make this field more robust, user friendly, and widespread.
Highlights
Analysis of intact proteins by native mass spectrometry has emerged as a powerful tool for obtaining insight into subunit diversity, post-translational modifications, stoichiometry, structural arrangement, stability, and overall architecture
An alternative strategy, which is the focus of this Perspective, is Mass spectrometry (MS)-based analysis of intact proteins, in which the full-length polypeptide is examined.[10−13] When native conditions are used for such an analysis, the intact protein is detected and the three-dimensional structure of the protein and its noncovalent interactions with protein partners and/or biomolecules are preserved.[10−12] This mode of analysis has been mainly the regime of native MS electrospray ionization (ESI) experiments, which has yielded unprecedented insights into the array of coexisting protein isoforms, the crosstalk between post-translational modifications (PTMs) and the structure and function of protein complexes ranging from ribosomes through viral capsids to membrane protein complexes
We propose that the MS analysis of intact proteins
Summary
Analysis of intact proteins by native mass spectrometry has emerged as a powerful tool for obtaining insight into subunit diversity, post-translational modifications, stoichiometry, structural arrangement, stability, and overall architecture. A recent MALDI-FTICR high-resolution imaging analysis of glioblastoma mouse brain tissue has identified a large number of intact proteins in the range of 4−15 kDa, including heavily modified histones.
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