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Titlebook: Deubiquitinases; Methods and Protocol Julie Maupin-Furlow,Mariola J. Edelmann Book 2023 The Editor(s) (if applicable) and The Author(s), un

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Identification of Deubiquitinase Substrates in , by Systematic Overexpression, in the cell, and second, redundancy between DUBs complicates loss of function screening approaches. Here, we describe a systematic overexpression approach that takes advantage of genome-wide resources available in . to overcome these challenges and identify DUB substrates in cells.
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Amit Patel,Andrew Crooks,Naoru Koizumilso outline the application of archaea in forming Ub/Ubl isopeptide linkages that include the covalent attachments of diverse archaeal and eukaryotic Ub/Ubls to target proteins. Archaea form these Ub/Ubl-linked protein conjugates in vivo, and the resulting products are found to serve as useful DUB substrates for in vitro assays.
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https://doi.org/10.1007/978-3-030-21406-7e pinpoint DUBs sufficient to counteract proteasomal degradation of these newly identified substrates. We apply this method to . egg extract but suggest that it can also be adapted to other cell lysates.
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Identification of Deubiquitinase Substrates in , Egg Extract,e pinpoint DUBs sufficient to counteract proteasomal degradation of these newly identified substrates. We apply this method to . egg extract but suggest that it can also be adapted to other cell lysates.
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