DICP OpenIR
Subject Area物理化学
A hydrophilic immobilized trypsin reactor with N-vinyl-2-pyrrolidinone modified polymer microparticles as matrix for highly efficient protein digestion with low peptide residue
Jiang, Hao1,2; Yuan, Huiming1; Liang, Yu1; Xia, Simin1,2; Zhao, Qun1,2; Wu, Qi1,2; Zhang, Lihua1; Liang, Zhen1; Zhang, Yukui1; Zhang LH(张丽华)
KeywordImmobilized Trypsin Reactors N-vinyl-2-pyrrolidinone Hydrophilicity Nano-rplc-esi-ms/ms Proteomics
Source PublicationJOURNAL OF CHROMATOGRAPHY A
2012-07-13
DOI10.1016/j.chroma.2012.03.014
Volume1246Pages:111-116
Indexed BySCI ; ISTP
SubtypeArticle
Department18
Funding Project1810
Contribution Rank1,1
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS KeywordTANDEM MASS-SPECTROMETRY ; ENZYME REACTORS ; CAPILLARY ; IDENTIFICATION ; SYSTEM ; MICROREACTOR ; PROTEOMICS ; MONOLITH ; ONLINE ; PROTEOLYSIS
AbstractIn this work, a novel kind of N-vinyl-2-pyrrolidinone (NVP) modified poly acrylic ester microspheres was prepared, followed by trypsin immobilization to prepare a hydrophilic immobilized enzyme reactor (IMER), to achieve highly efficient protein digestion with low peptide residue. The nonspecific adsorption of peptides on such an IMER was evaluated by the in sequence digestion of bovine serum albumin (BSA) and myoglobin. Without NVP modification, both proteins could be identified after digestion by a 5 cm-length IMER, but 18 peptides of BSA were found in the digests of myoglobin caused by the nonspecific adsorption of the matrix. With NVP modification, the hydrophilicity of IMER was greatly improved, resulting in not only the sequence coverage of myoglobin increased from 63% to 73%, but also no residual peptides from BSA observed in myoglobin digests. Although the sequence coverages of proteins obtained by the IMER were comparable to those obtained by in-solution digestion, the digestion time was shortened from 24 h to 1 min. By such an IMER, a protein mixture, containing BSA, myoglobin, and cytochrome c (100, 1 and 0.01 mu g/mL, respectively), was digested, and all proteins were unambiguously identified with improved sequence coverages than that achieved by in-solution digestion. Furthermore, the hydrophilic IMER was also off-line coupled to nano-RPLC-ESI-MS/MS for the analysis of proteins extracted from yeast. After 1.5 min digestion, 271 protein groups with at least 2 distinct peptides were identified, much more than those obtained by 24 h in-solution digestion (192 protein groups), indicating the great potential of such an IMER for proteome analysis. (C) 2012 Elsevier B.V. All rights reserved.
Language英语
WOS IDWOS:000306246000018
Citation statistics
Cited Times:23[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/118205
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhang LH(张丽华)
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
Recommended Citation
GB/T 7714
Jiang, Hao,Yuan, Huiming,Liang, Yu,et al. A hydrophilic immobilized trypsin reactor with N-vinyl-2-pyrrolidinone modified polymer microparticles as matrix for highly efficient protein digestion with low peptide residue[J]. JOURNAL OF CHROMATOGRAPHY A,2012,1246:111-116.
APA Jiang, Hao.,Yuan, Huiming.,Liang, Yu.,Xia, Simin.,Zhao, Qun.,...&张丽华.(2012).A hydrophilic immobilized trypsin reactor with N-vinyl-2-pyrrolidinone modified polymer microparticles as matrix for highly efficient protein digestion with low peptide residue.JOURNAL OF CHROMATOGRAPHY A,1246,111-116.
MLA Jiang, Hao,et al."A hydrophilic immobilized trypsin reactor with N-vinyl-2-pyrrolidinone modified polymer microparticles as matrix for highly efficient protein digestion with low peptide residue".JOURNAL OF CHROMATOGRAPHY A 1246(2012):111-116.
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