title: A novel method to isolate protein N-terminal peptides from proteome samples using sulfydryl tagging and gold-nanoparticle-based depletion.
abstract: A novel method to isolate global N-termini using sulfydryl tagging and gold-nanoparticle-based depletion (STagAu method) is presented. The N-terminal and lysine amino groups were first completely dimethylated at the protein level, after which the proteins were digested. The newly generated internal peptides were tagged with sulfydryl by Traut's reagent through digested N-terminal amines in yields of 96%. The resulting sulfydryl peptides were depleted through binding onto nano gold composite materials. The Au-S bond is stable and widely used in materials science. Nano gold composite materials showed nearly complete depletion of sulfydryl peptides. A set of the acetylated and dimethylated N-terminal peptides were analyzed by liquid chromatography-tandem mass spectrometry. This method was demonstrated to be an efficient N-terminus enrichment method because of the use of an effective derivatization reaction, in combination with robust and relative easy to implement Au-S coupling. We identified 632 N-terminal peptides from 386 proteins in a mouse liver sample. The STagAu approach presented is therefore a facile and efficient method for mass-spectrometry-based analysis of proteome N-termini or protease-generated cleavage products.
authors.affiliation: Department of Chemistry, Fudan University, Shanghai, 200433, China.|Institute of Biomedical Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Lanting
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Chemistry, Fudan University, Shanghai, 200433, China.|Institute of Biomedical Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Runqing
authors.initials: R
authors.lastname: Wu
authors.affiliation: Department of Chemistry, Fudan University, Shanghai, 200433, China.|Institute of Biomedical Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Guoquan
authors.initials: G
authors.lastname: Yan
authors.affiliation: Department of Chemistry, Fudan University, Shanghai, 200433, China.|Institute of Biomedical Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Mingxia
authors.initials: M
authors.lastname: Gao
authors.affiliation: Department of Chemistry, Fudan University, Shanghai, 200433, China.|Institute of Biomedical Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Chunhui
authors.initials: C
authors.lastname: Deng
authors.affiliation: Department of Chemistry, Fudan University, Shanghai, 200433, China. xmzhang@fudan.edu.cn.|Institute of Biomedical Sciences, Fudan University, Shanghai, 200433, China. xmzhang@fudan.edu.cn.
authors.forename: Xiangmin
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000818:Animals; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D008099:Liver / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry*; D051379:Mice; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D020543:Proteome / Q000737:chemistry*; D040901:Proteomics; D053719:Tandem Mass Spectrometry
chemical_list: D010455:Peptides; D011506:Proteins; D020543:Proteome; D006046:Gold
keywords: Gold nanoparticle; N-Termini; Posttranslational modification; Proteomics; Traut’s reagent