DiLeu Isobaric Labeling Coupled with Limited Proteolysis Mass Spectrometry for High-Throughput Profiling of Protein Structural Changes in Alzheimer’s Disease
Year: 2023
Type: article
Abstract: High-throughput quantitative analysis of protein conformational changes has a profound impact on our understanding of the pathological mechanisms of Alzheimer’s disease (AD). To establish an effective workflow enabling quantitative analysis of changes in protein conformation within multiple samples simultaneously, here we report the combination of N,N -dimethyl leucine (DiLeu) isobaric tag labeling with limited proteolysis mass spectrometry (DiLeu-LiP-MS) for high-throughput structural protein quantitation in serum samples collected from AD patients and control donors. Twenty-three proteins were discovered to undergo structural changes, mapping to 35 unique conformotypic peptides with significant changes between the AD group and the control group. Seven out of 23 proteins, including CO3, CO9, C4BPA, APOA1, APOA4, C1R, and APOA, exhibited a potential correlation with AD. Moreover, we found that complement proteins (e.g., CO3, CO9, and C4BPA) related to AD exhibited elevated levels in the AD group compared to those in the control group. These results provide evidence that the established DiLeu-LiP-MS method can be used for high-throughput structural protein quantitation, which also showed great potential in achieving large-scale and in-depth quantitative analysis of protein conformational changes in other biological systems.
Source: Analytical Chemistry
Authors: Haiyan Lu, Bin Wang, Yuan Liu, Danqing Wang, Lauren Fields, Hua Zhang, Miyang Li, Xudong Shi, Henrik Zetterberg, Lingjun Li
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: green
Funders: Alzheimer's Association, Alzheimer's Drug Discovery Foundation, Familjen Erling-Perssons Stiftelse, Stiftelsen för Gamla Tjänarinnor, EU Joint Programme – Neurodegenerative Disease Research, University College London, European Commission, Hjärnfonden, Vetenskapsrådet, UK Dementia Research Institute, Olav Thon Stiftelsen, National Institutes of Health, National Institute on Aging, National Institute of General Medical Sciences, National Institute of Diabetes and Digestive and Kidney Diseases, National Center for Research Resources, H2020 European Research Council, H2020 Marie Skłodowska-Curie Actions, American Society for Mass Spectrometry
Awards: G1057200632, G1088883541, G150733400, G1551449788, G1588116593, Acquisition of a High-Field Dual Source FTICR-MS for Pharmaceutical Research, G176366027, G1802301745, G1833008592, G201353144, G2057385626, G2118924489, G237833635, G2388130288, G2543812357, G2560624041, G2799940023, G292763017, G3055837027, G3241251457, G3370889232, G3418984644, G3903502641, G3914690605, G4234328627, G4260843567, G432431387, G434337789, G4377393254, G5040492720, G506008078, G5111822078, G516766298, Multi-omics Interdisciplinary Research Integration to Address DEmentia diagnosis, G6289970089, Multi-omics Interdisciplinary Research Integration to Address DEmentia diagnosis, G691317373, Pathways to Alzheimer's disease, Acquisition of a High Resolution High Speed MALDI Mass Spectrometer for Biomedical Research at UW-Madison, G712881263, Acquisition of a Dual-Source, High-Performance, Ion Mobility, Quadrupole Time-of-Flight Mass Spectrometry System for Biomedical Research at UW-Madison, G822836364, What determines amyloid beta aggregation and toxicity in Alzheimer’s disease?, G9000023674, G9000068044, G9000197343, G9000228399, G9000231773, Probing Protein Structural Changes in Alzheimers Disease
OpenAlex ID: W4380291971