Lee, Seonjeong Ju, Shinyeong Kim, Seok Jin Choi, Jin-Oh Kim, Kihyun Kim, Darae Jeon, Eun-Seok Lee, Cheolju 2024-01-19T13:32:35Z 2024-01-19T13:32:35Z 2022-01-10 2021-10-26 0003-2700 https://pubs.kist.re.kr/handle/201004/116250 The mass spectrometry-based analysis of protein post-translational modifications requires large amounts of sample, complicating the analysis of samples with limited amounts of proteins such as clinical biopsies. Here, we present a tip-based N-terminal analysis method, tipNrich. The entire procedure is processed in a single pipette tip to minimize sample loss, which is so highly optimized to analyze small amounts of proteins, even femtomole-scale of a single protein. With tipNrich, we investigated various single proteins purified from different organisms using a low-resolution mass spectrometer and identified several N-terminal peptides with different Nt-modifications such as ragged N-termini. Furthermore, we applied matrix-assisted laser desorption ionization time-of-flight mass spectrometry to our method for shortening the analysis time. Moreover, we showed that our method could be utilized in disease diagnosis as exemplified by the characterization of wild-type transthyretin amyloidosis patients compared to the healthy individuals based on N-terminome profiling. In summary, tipNrich will satisfy the need of identifying N-terminal peptides even with highly scarce amounts of proteins and of having faster processing time to check the quality of protein products or to characterize N-terminal proteoform-related diseases. English AMER CHEMICAL SOC CLEAVAGE SITES HUMAN PLASMA TRANSTHYRETIN PROTEINS PEPTIDES MALDI TOOL IDENTIFICATION GLYCOSYLATION PURIFICATION tipNrich: A Tip-Based N-Terminal Proteome Enrichment Method Article 10.1021/acs.analchem.1c01722 1 ANALYTICAL CHEMISTRY, v.93, no.42, pp.14088 - 14098 ANALYTICAL CHEMISTRY 93 42 14088 14098 scie scopus 000711718700010 2-s2.0-85117453099 Chemistry, Analytical Chemistry Article CLEAVAGE SITES HUMAN PLASMA TRANSTHYRETIN PROTEINS PEPTIDES MALDI TOOL IDENTIFICATION GLYCOSYLATION PURIFICATION