title: Hydrophobic Tagging-Assisted N-Termini Enrichment for In-Depth N-Terminome Analysis.
abstract: The analysis of protein N-termini is of great importance for understanding the protein function and elucidating the proteolytic processing. Herein, we develop a negative enrichment strategy, termed as hydrophobic tagging-assisted N-termini enrichment (HYTANE) to achieve a global N-terminome analysis. The HYTANE strategy showed a high efficiency in hydrophobic tagging and C18 material-assisted depletion using bovine serum albumin (BSA) as the sample. This strategy was applied to N-termini profiling from S. cerevisiae cell lysates and enabled the identification of 1096 protein N-termini, representing the largest N-terminome data set of S. cerevisiae. The identified N-terminal peptides accounted for 99% of all identified peptides, and no deficiency in acidic, histidine (His)-containing, and His-free N-terminal peptides was observed. The presented HYTANE strategy is therefore a highly selective, efficient, and unbiased strategy for the large scale N-terminome analysis. Furthermore, using the HYTANE strategy, we identified 329 cleavage sites and 291 substrates of caspases in Jurkat cells, demonstrating the great promise of HYTANE strategy for protease research. Data are available via ProteomeXchange with identifier PXD004690.
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian, Liaoning 116023, China.|University of the Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Lingfan
authors.initials: L
authors.lastname: Chen
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian, Liaoning 116023, China.
authors.forename: Yichu
authors.initials: Y
authors.lastname: Shan
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian, Liaoning 116023, China.|University of the Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Yejing
authors.initials: Y
authors.lastname: Weng
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian, Liaoning 116023, China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Sui
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian, Liaoning 116023, China.
authors.forename: Xiaodan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian, Liaoning 116023, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Liang
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian, Liaoning 116023, China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian, Liaoning 116023, China.
authors.forename: Yukui
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D020169:Caspases / Q000378:metabolism; D002417:Cattle; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D019169:Jurkat Cells; D010446:Peptide Fragments / Q000032:analysis* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000032:analysis* / Q000378:metabolism*
chemical_list: D010446:Peptide Fragments; D012710:Serum Albumin, Bovine; D020169:Caspases