Description
The recent development of high-resolution mass spectrometry enables to study proteomes at an unprecedented depth even at single cell level. Importantly the developments with proteomic sample preparation methods, nano liquid chromatography, sensitivity of mass spectrometry instrumentation it is now possible to identify and quantify >12,000 proteins per sample which equates to the coverage that is often achieved with transcriptomics. While these efforts led to achieve high-sensitivity and coverage the throughput has become a major limitation in MS-based proteomics. The recent Thermo “Orbitrap Astral Mass spectrometer” enables to measure >200 proteomes per day still achieving highest possible coverage and sensitivity. I will discuss specific use cases of Orbitrap Astral and how this technology is transformative in studying Organelle and Spatial proteomes and post-translational modifications such as protein phosphorylation and Ubiquitylation in understanding neurodegenerative diseases such as Parkinson’s and Amyotrophic lateral sclerosis. I will specifically highlight the development of rapid immunoprecipitation of Organelle (Golgi-IP, Lysosome-IP and mitochondria-IP) that enables to study molecular content of intact Organelles at high-resolution and study how this is linked to disease processes such as Parkinson’s. Current methods enable the analysis of protein, metabolite, and lipid content to be assessed in depth using high-resolution mass spectrometry analysis. However, due to the low levels of proteins obtained from organelle IPs typically under 5 to 10 microgram amounts it has thus far not been possible to undertake high resolution profiling of post translational modifications (PTMs) such as protein phosphorylation and Ubiquitylation in these fractions. We developed sensitive workflows to define PTM landscape of organelles using Orbitrap Astral. I will also discuss our efforts in leveraging Orbitrap platforms in development of multiplexed targeted PRM assays to assess LRRK2 kinase (implicated in Parkinson’s disease) activity in cells, tissues and body fluids. We believe our workflows could open new avenues for studying Organelle specific phosphorylation and ubiquitylation signalling in health and disease.| Period | 1 Oct 2024 → 4 Oct 2024 |
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| Degree of Recognition | International |