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Total Chemical Synthesis of ISGylated-Ubiquitin Hybrid Chain Assisted by Acetamidomethyl Derivatives with Dual Functions
journal contribution
posted on 2020-02-21, 21:32 authored by Emad Eid, Gábor N. Boross, Hao Sun, Muna Msallam, Sumeet K. Singh, Ashraf BrikInterferon-stimulated
gene 15 (ISG15) is a member of the ubiquitin-like
modifiers (ULM) family, which adopts a β-grasp fold domain(s)
similar to ubiquitin (Ub) with only minor sequence homology. ISG15
consists of two Ub-like domains and aids the immune system in neutralizing
infections by numerous pathogens and plays an important role in defending
cells against many viruses including influenza A. Recently, Ub was
found to be a substrate for ISG15, which can be ISGylated on Lys29
and Lys48, while the former is more dominant. The discovery of such
hybrid ISG15-Ub chains brought forward various fundamental questions
regarding the nature and effect of this conjugation. To further investigate
the role of hybrid ISG15-Ub chains, the pure homogeneous material
of these chains is needed in workable quantities. By applying advanced
chemical strategies for protein synthesis, we report the total chemical
synthesis of a 231-residue ISG15-Lys29-Ub hybrid chain. During the
synthesis we encountered insoluble peptide fragments, and therefore
we developed a new reversible Acm based solubilizing tag to efficiently
tackle this hurdle. This new Acm tag was compared with the known Arg
based Acm solubilizing tag and was found to be more reliable in terms
of incorporation and efficiency as demonstrated in the synthesis of
the native ISG15-Ub hybrid chain.
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Keywords
chemical synthesisAcm solubilizing tagISGylated-Ubiquitin Hybrid Chain AssistedISG 15-UbDual Functions Interferon-stimulated gene 15231- residue ISG 15-LysAcetamidomethyl Derivativesubiquitin-like modifiersneutralizing infectionssequence homologyLys 29Ub-like domainsTotal Chemical Synthesischemical strategiespeptide fragmentsprotein synthesisAcm tagsolubilizing tagISG 15-Ub chainsULMISG 15
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