SEC-SWATH-MS

Explain Like I'm Five | ELI5

To my friends and family not in the biopharma industry:

• Envision possessing a sizable container of LEGOs, with which you can construct an array of objects such as houses, automobiles, and spacecrafts.
• Now, envisage that within your corporeal form, there exist minuscule building blocks termed proteins. These proteins can coalesce to create more substantial entities designated protein complexes, which execute crucial functions to maintain the operation of your body.
• Researchers wanted to observe the alterations of these protein complexes during cellular growth and division. Thus, they employed a specialized instrument denominated SEC-SWATH-MS to examine the proteins and protein complexes in cells. They discovered that during cellular growth and division, the protein complexes undergo significant modifications. Additionally, they developed a computational program to facilitate other scientists in analyzing the information they obtained.

Is that not remarkable? Scientists can utilize specialized instruments to acquire knowledge about the building blocks within our bodies!

A Global Screen for Assembly State Changes of the Mitotic Proteome by SEC-SWATH-MS

Moritz Heusel et al. describe an integrated technique to quantify cell-state-specific changes in the physical arrangement of protein complexes concurrently for thousands of proteins and hundreds of complexes. The technique was applied to a comparison of human cells in interphase and mitosis, providing a systematic overview of mitotic proteome reorganization.

Facts

?? The technique used is called SEC-SWATH-MS, which combines size exclusion chromatography with SWATH mass spectrometry.

?? The study was performed on human cells in interphase and mitosis.

?? The results provide a systematic overview of mitotic proteome reorganization.

?? The study recalls key hallmarks of mitotic complex remodeling.

?? The study introduces SECexplorer-cc, an interactive exploration tool to support the interpretation of quantitative SEC-SWATH-MS datasets.

Notes:

https://sec-explorer.shinyapps.io/hela_cellcycle/

Size Exclusion Chromatography Column

• Sepax Technologies, Inc.?SRT-C SEC-500, 5um, 500 A 21.2 x 300 mm
• Part Number:?235500-21230

Flow Rate

• 2 ml/min flow rate on ice (0–4°C)

Mobile Phase

• 50 mM HEPES pH 7.5, 150 mM NaCl

Citation

Heusel, Moritz, et al. "A global screen for assembly state changes of the mitotic proteome by SEC-SWATH-MS." Cell systems 10.2 (2020): 133-155.

DOI: 10.1016/j.cels.2020.01.001

Authors

Moritz Heusel ,? Max Frank , Mario K?hler , Sabine Amon , Fabian Frommelt , George Rosenberger , Isabell Bludau, PhD , Simran A. , Monika I. Linder , Yansheng Liu , Ben Collins , Matthias Gstaiger , Ulrike Kutay, Ruedi Aebersold ?

Author Affiliations

1. Institute of Molecular Systems Biology, 瑞士苏黎世联邦理工学院 , Zurich, Switzerland
2. Institute of Biochemistry, ETH Zurich, Zurich, Switzerland
3. Department of Biology, Institute of Molecular Systems Biology, ETH Zurich, Zurich, Switzerland
4. Functional Genomics Center Zurich (FGCZ), University of Zurich/ETH Zurich, Zurich, Switzerland
5. PhD Program in Systems Biology, University of Zurich and ETH Zurich, Zurich, Switzerland
6. Faculty of Science, University of Zurich, Zurich, Switzerland
7. Competence Center Personalized Medicine UZH/ETH (CCPM), Universit?tsspital Zürich and University of Zurich/ETH Zurich, Zurich, Switzerland
8. PhD Program in Molecular and Translational Biomedicine (MTB), University of Zurich and ETH Zurich, Zurich, Switzerland

Funding Institutions

Swiss National Science Foundation SNSF (SNSF), European Research Council (ERC) , Swiss Initiative in Systems Biology (SystemsX.ch), The National Institutes of Health (NIH)