Dataset Information

Extending the coverage of Lys-C/trypsin-based bottom-up proteomics using cysteine S-aminoethylation

ABSTRACT: In this study, S-aminoethylation of Cys (AE-Cys) was carried out with 2-bromoethylamine to form pseudo-Lys, which was then digested with Lys-C or trypsin to improve proteome coverage in bottom-up proteomics. A model study with bovine serum albumin showed that the C-terminal side of Cys was successfully cleaved by AE-Cys followed by digestion with Lys-C. The frequency of side reactions to amino acids other than Cys was not as high as that of carbamidomethylation of Cys with iodoacetamide. Proteomic analysis of A549 cell extracts using AE-Cys identified more protein groups, especially membrane proteins, in a data-dependent acquisition mode. In addition, label-free quantification of mouse non-small cell lung cancer (NSCLC) tissue in data-independent acquisition mode showed improved NSCLC pathway coverage and higher reproducibility with AE-Cys. Furthermore, we successfully identified peptides containing the [T790M] mutation site of EGFR by AE-Cys, which has not been reported by conventional bottom-up methods despite its importance as one of the most important lung cancer-related mutation sites.

ORGANISM(S): Homo Sapiens (human) Bos Taurus

SUBMITTER: Yasushi Ishihama

PROVIDER: PXD045433 | JPOST Repository | Wed Sep 27 00:00:00 BST 2023

REPOSITORIES: jPOST

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Dataset's files

Source:

			Action	DRS
	A549_BEA_LysC.zip	Other
	A549_BEA_LysCTrypsin.zip	Other
	A549_BEA_LysCTrypsin_result.zip	Other
	A549_BEA_LysC_result.zip	Other
	A549_IAA_LysC.zip	Other

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Publications

Extending the Coverage of Lys-C/Trypsin-Based Bottom-up Proteomics by Cysteine S-Aminoethylation.

Tomioka Ryota R Tomioka Ayana A Ogata Kosuke K Chan Hsin-Ju HJ Chen Li-Yu LY Guzman Ulises H UH Xuan Yue Y Olsen Jesper V JV Chen Yu-Ju YJ Ishihama Yasushi Y

Journal of the American Society for Mass Spectrometry 20240129 2

To improve the coverage in bottom-up proteomics, S-aminoethylation of cysteine residues (AE-Cys) was carried out with 2-bromoethylamine, followed by cleavage with lysyl endopeptidase (Lys-C) or Lys-C/trypsin. A model study with bovine serum albumin showed that the C-terminal side of AE-Cys was successfully cleaved by Lys-C. The frequency of side reactions at amino acids other than Cys was less than that in the case of carbamidomethylation of Cys with iodoacetamide. Proteomic analysis of A549 cel ...[more]

PMID: 38287222

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