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• W1983223542 abstract "Post-translational modification of proteins via ubiquitination plays a crucial role in numerous vital functions of the cell. Polyubiquitination is one of the key regulatory processes involved in regulation of mitotic progression. Here we describe a differential proteomic screen dedicated to identification of novel proteins ubiquitinated upon mitotic exit in cell-free extract of Xenopus laevis embryo. Mutated recombinant His6-tagged ubiquitin (UbiK48R) was added to mitotic extract from which we purified conjugated proteins, as well as associated proteins in nondenaturing conditions by cobalt affinity chromatography. Proteins eluted from UbiK48R supplemented and control extracts were compared by LC-MS/MS analysis after monodimensional SDS-PAGE. A total of 144 proteins potentially ubiquitinated or associated with them were identified. Forty-one percent of these proteins were shown to be involved in ubiquitination and/or proteasomal degradation pathway confirming the specificity of the screen. Twelve proteins, among them ubiquitin itself, were shown to carry a “GG” or “LRGG” remnant tag indicating their direct ubiquitination. Interestingly, sequence analysis of ubiquitinated substrates carrying these tags indicated that in Xenopus cell-free embryo extract supplemented with UbiK48R the majority of polyubiquitination occurred through lysine-11 specific ubiquitin chain polymerization. The potential interest in this atypical form of ubiquitination as well as usefulness of our method in analyzing atypical polyubiquitin species is discussed." @default.
• W1983223542 created "2016-06-24" @default.
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• W1983223542 date "2008-09-30" @default.
• W1983223542 modified "2023-10-02" @default.
• W1983223542 title "Differential Proteomic Screen To Evidence Proteins Ubiquitinated upon Mitotic Exit in Cell-Free Extract of <i>Xenopus laevis</i> Embryos" @default.
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• W1983223542 doi "https://doi.org/10.1021/pr800250x" @default.
• W1983223542 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/18823142" @default.
• W1983223542 hasPublicationYear "2008" @default.
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