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In Planta Chemical Cross-linking and Mass Spectrometry Analysis of Protein Structure and Interaction in Arabidopsis

Authors Zhu, Xinliang HKUST affiliated (currently or previously)
Yu, Fengchao HKUST affiliated (currently or previously)
Yang, Zhu HKUST affiliated (currently or previously)
Liu, Shichang HKUST affiliated (currently or previously)
Dai, Chen
Lu, Xiaoyun
Liu, Chenyu HKUST affiliated (currently or previously).
Yu, Weichuan View this author's profile
Li, Ning View this author's profile
Issue Date 2016
Source Proteomics , v. 16, (13), July 2016, p. 1915-1927
Summary Site-specific chemical cross-linking in combination with mass spectrometry analysis (CX-MS) has emerged as a powerful proteomic approach for studying the three-dimensional structure of protein complexes and in mapping protein-protein interactions (PPI). Building on the success of mass spectrometry analysis of in vitro cross-linked proteins, which has been widely used to investigate specific interactions of bait proteins and their targets in various organisms, we report a workflow for in vivo chemical cross-linking and mass spectrometry analysis in a multicellular eukaryote. This approach optimizes the in vivo protein cross-linking conditions in Arabidopsis thaliana, establishes a MudPIT procedure for the enrichment of cross-linked peptides, and develops an integrated software program, ECL, to identify the MS spectra of in planta chemical cross-linked peptides. In total, two pairs of in vivo cross-linked peptides of high-confidence have been identified from two independent biological replicates. This work demarks the beginning of an alternative proteomic approach in the study of in vivo protein tertiary structure and PPIs in multicellular eukaryotes.
ISSN 1615-9853
Language English
Format Article
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