Ju, Shinyeong Kwon, Yumi Kim, Jeong-Mok Park, Daechan Lee, Seonjeong Lee, Jin-Won Hwang, Cheol-Sang Lee, Cheolju 2024-01-19T17:32:43Z 2024-01-19T17:32:43Z 2021-09-05 2020-05-05 0003-2700 https://pubs.kist.re.kr/handle/201004/118631 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. English AMER CHEMICAL SOC PROTEOLYTIC EVENTS PEPTIDES PROTEINS TERMINOMICS INHIBITION ACTIVATION APOPTOSIS PROJECT TAILS iNrich, Rapid and Robust Method to Enrich N-Terminal Proteome in a Highly Multiplexed Platform Article 10.1021/acs.analchem.9b05653 1 ANALYTICAL CHEMISTRY, v.92, no.9, pp.6462 - 6469 ANALYTICAL CHEMISTRY 92 9 6462 6469 scie scopus 000530658600040 2-s2.0-85084835014 Chemistry, Analytical Chemistry Article PROTEOLYTIC EVENTS PEPTIDES PROTEINS TERMINOMICS INHIBITION ACTIVATION APOPTOSIS PROJECT TAILS