MassIVE MSV000089163

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Spatially resolved top-down proteomics of tissue sections based on a microfluidic nanodroplet sample preparation platform

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Description

We demonstrate spatially resolved top-down proteomics (TDP) for proteoform identification and quantification from small sections of rat brain. We used the nanoPOTS (nanodroplet Processing in One pot for Trace Samples) platform, further improving the sample preparation by adding benzonase in the extraction buffer. We increased identified proteoforms from 493 to 700 in 200 cultured cells, with the largest improvements among nuclear proteins. We then performed nanoPOTS TDP on 100,000 um2 (around 200 cells) laser capture microdissectioned rat brain cortex/hypothalamus tissues. The extracted proteins were directly subjected to top down analysis without enzymatic digestion. Data was searched with TopPIC and TDPortal. [doi:10.25345/C5CC0TX6N] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Top-down proteomics ; nanoPOTS ; proteoform ; spatial proteomics ; imaging

Contact

Principal Investigators:
(in alphabetical order) 		Mowei Zhou, Pacific Northwest National Laboratory, United States
Submitting User: alchemistmatt

Publications

Liao YC, Fulcher JM, Degnan DJ, Williams SM, Bramer LM, Veli?kovi? D, Zemaitis KJ, Veli?kovi? M, Sontag RL, Moore RJ, Paša-Toli? L, Zhu Y, Zhou M.
Spatially Resolved Top-Down Proteomics of Tissue Sections Based on a Microfluidic Nanodroplet Sample Preparation Platform.
Mol Cell Proteomics. 2023 Feb;22(2):100491. Epub 2023 Jan 2.

Yen-Chen Liao, James Fulcher, David Degnan, Sarah Williams, Lisa Bramer, Dušan Veli?kovi?, Kevin Zemaitis, Marija Veli?kovi?, Ryan Sontag, Ronald Moore, Ljiljana Paša-Toli?, Ying Zhu, Mowei Zhou.
Spatially resolved top-down proteomics of tissue sections based on a microfluidic nanodroplet sample preparation platform.
chemRxiv. Epub 2022 Jun 28. doi: https://doi.org/10.26434/chemrxiv-2022-pcn8c.

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