iNrich, Rapid and Robust Method to Enrich N-Terminal Proteome in a Highly Multiplexed Platform
SCIE
SCOPUS
- Title
- iNrich, Rapid and Robust Method to Enrich N-Terminal Proteome in a Highly Multiplexed Platform
- Authors
- Ju, Shinyeong; Kwon, Yumi; Kim, Jeong-Mok; Park, Daechan; Lee, Seonjeong; Lee, Jin-Won; Hwang, Cheol-Sang; Lee, Cheolju
- Date Issued
- 2020-05
- Publisher
- AMER CHEMICAL SOC
- Abstract
- The field of terminal proteomics is limited in that it is optimized for large-scale analysis via multistep processes involving liquid chromatography. Here, we present an integrated N-terminal peptide enrichment method (iNrich) that can handle as little as 25 mu g of cell lysate via a single-stage encapsulated solid-phase extraction column. iNrich enables simple, rapid, and reproducible sample processing, treatment of a wide range of protein amounts (25 mu g similar to 1 mg), multiplexed parallel sample preparation, and in-stage sample prefractionation using a mixed-anion-exchange filter. We identified similar to 5000 N-terminal peptides (Nt-peptides) from only 100 mu g of human cell lysate including Nt-formyl peptides. Multiplexed sample preparation facilitated quantitative and robust enrichment of N-terminome iNrich with dozens of samples simultaneously. We further developed the method to incorporate isobaric tags such as a tandem mass tag (TMT) and used it to discover novel peptides during ER stress analysis. The iNrich facilitated high-throughput N-terminomics and degradomics at a low cost using commercially available reagents and apparatus, without requiring arduous procedures.
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/104830
- DOI
- 10.1021/acs.analchem.9b05653
- ISSN
- 0003-2700
- Article Type
- Article
- Citation
- ANALYTICAL CHEMISTRY, vol. 92, no. 9, page. 6462 - 6469, 2020-05
- Files in This Item:
- There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.