Mass spectrometric insights into protein aggregation

Abstract

Protein aggregation is now recognized as a generic and significant component of the protein energy landscape. Occurring through a complex and dynamic pathway of structural interconversion, the assembly of misfolded proteins to form soluble oligomers and insoluble aggregates remains a challenging topic of study, both in vitro and in vivo. Since the etiology of numerous human diseases has been associated with protein aggregation, and it has become a field of increasing importance in the biopharmaceutical industry, the biophysical characterization of protein misfolded states and their aggregation mechanisms continues to receive increased attention. Mass spectrometry (MS) has firmly established itself as a powerful analytical tool capable of both detection and characterization of proteins at all levels of structure. Given inherent advantages of biological MS, including high sensitivity, rapid timescales of analysis, and the ability to distinguish individual components from complex mixtures with unrivalled specificity, it has found widespread use in the study of protein aggregation, importantly, where traditional structural biology approaches are often not amenable. The present review aims to provide a brief overview of selected MS-based approaches that can provide a range of biophysical descriptors associated with protein conformation and the aggregation pathway. Recent examples highlight where this technology has provided unique structural and mechanistic understanding of protein aggregation.

Nanoplastic Stimulates the Amyloidogenesis of Parkinson's Alpha‐Synuclein NACore

“…Methodologically, this marked the first use of high-resolution MS for the study of amyloid protein-nanoparticle interactions, expanding the literature where MS has been employed for characterizing the early aggregation of amyloid proteins, the catalytic cross-talk between Alzheimer's A𝛽 [25][26][27][28][29][30][31][32][33][34][35] and amyotrophic lateral sclerosis's TDP43 307-319 , the screening of small-molecular inhibitors against the aggregation of A𝛽 40 and IAPP, as well as the binding of metal ions with 𝛼S in aggregation. [42][43][44][45][46][47][48][49][90][91][92] As demonstrated by the present study, ESI-TOF-MS was able to resolve the rate of production and sub-populations of the toxic NACore oligomeric species that would otherwise be indistinguishable by conventional techniques. As assessment of amyloid protein-nanoparticle interactions is central to the development of amyloid nanomedicine and elucidating the environment-human health axis, this study opened the door to a range of new applications exploiting the non-invasive, high-throughput, and highresolution MS techniques.…”

“…Owing to their distinct advantages of high throughput, small sample volume, and rapid sampling without ensemble averaging, MS in its various forms has recently found a niche in probing the aggregation of amyloid proteins of amyloid beta (A𝛽), 𝛼Bcrystallin, and human islet amyloid polypeptide (IAPP), the inhibitory potential of small molecules against amyloidosis, as well as metal binding with 𝛼S in aggregation. [42][43][44][45][46][47][48][49] However, no research to date has employed MS for the examination of amyloidosis with nanomaterials, an area pertinent to the development of brain-targeting nanomedicine and to the elucidation of environmental health and safety against discharged nanomaterials and nano-pollutants. [4,[50][51] Biophysically, amyloid aggregation occurs spontaneously by the self-assembly of proteins to evolve from amphiphilic monomers to water-soluble oligomers and, eventually, to colloid-like amyloid fibrils.…”

Probing protein aggregation through spectroscopic insights and multimodal approaches: A comprehensive review for counteracting neurodegenerative disorders

“…It provides valuable information about the initiation of amorphous aggregation and, more specifically, the formation of amyloid fibrils. This includes the characterization of monomers and oligomers, the study of aggregation mechanisms, and the screening of inhibitors [ 108 ]. Furthermore, this technique is used for the investigation of proteins, peptides and oligomers [ 109 ].…”

Stabilization challenges and aggregation in protein-based therapeutics in the pharmaceutical industry

“…While partially unfolded proteins may possess the ability to refold under specic conditions, the thermodynamics and kinetics generally favour the aggregation process over refolding. [92][93][94] Another mechanism is that the native protein monomers can aggregate by adhering to pre-existing protein oligomers, contaminants or to the surfaces of containers. This adherence can initiate a nucleation process in which the aggregates rapidly grow in size.…”