Modifying the Vicinity of the Isopeptide Bond To Reveal Differential Behavior of Ubiquitin Chains with Interacting Proteins: Organic Chemistry Applied to Synthetic Proteins

Haj-Yahya, Najat; Haj‐Yahya, Mahmood; Castañeda, Carlos; Spasser, Liat; Hemantha, Hosahalli P.; Jbara, Muhammad; Penner, Marlin; Ciechanover, Aaron; Fushman, David; Brik, Ashraf

doi:10.1002/anie.201306118

Cited by 27 publications

(25 citation statements)

References 40 publications

(26 reference statements)

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“…Examining the reported cases for the expression of ISG15, it is established that Cys78 significantly contributes to the precipitation of the full ISG15 protein. 7,25 Subsequently, we decided to introduce the Cys78Ser mutation in order to avoid the formation of the dimer form of the protein. This mutation is located in the loop region, connecting the two Ub like domains of ISG15; thus no impact on the biological activity is anticipated.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Total Chemical Synthesis of ISGylated-Ubiquitin Hybrid Chain Assisted by Acetamidomethyl Derivatives with Dual Functions

et al. 2020

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Interferon-stimulated gene 15 (ISG15) is a member of the ubiquitin-like modifiers (ULM) family, which adopts a βgrasp 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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Section: ■ Results and Discussionmentioning

confidence: 99%

Total Chemical Synthesis of ISGylated-Ubiquitin Hybrid Chain Assisted by Acetamidomethyl Derivatives with Dual Functions

et al. 2020

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“…Lys(Cys) peptides have also been used successfully by other groups for accessing to ubiquitinated proteins. 39,40 An alternative would be to use a target peptide featuring a g- 41,42 or d-mercaptolysine 14,15,18 residue. However, the access to a native SUMO-1 conjugate using this latter strategy would require protecting the Cys51 thiol group during the desulfurization of the mercaptolysine residue.…”

Section: Resultsmentioning

confidence: 99%

A novel PEG-based solid support enables the synthesis of >50 amino-acid peptide thioesters and the total synthesis of a functional SUMO-1 peptide conjugate

et al. 2014

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“…Using a method similar to that for K 27 ‐isoUb ( 2 ), 9 was readily synthesized through Fmoc SPPS in 19.5 % yield of isolated product. Peptide 10 was similarly prepared through Fmoc SPPS in 26 % yield of isolated product.…”

Section: Methodsmentioning

confidence: 99%

Practical Chemical Synthesis of Atypical Ubiquitin Chains by Using an Isopeptide‐Linked Ub Isomer

Tang

Liang

et al. 2017

Angewandte Chemie

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Chemical ubiquitination is an effective approach for accessing structurally defined, atypical ubiquitin (Ub) chains that are difficult to prepare by other techniques.H erein, we describe as trategy that uses ar eadily accessible premade isopeptide-linked 76-mer (isoUb), whichh as an N-terminal Cys and aC -terminal hydrazide,a st he key building blockt o assemble atypical Ub chains in am odular fashion. This method avoids the use of auxiliary-modified Lysa nd instead employs the canonical and therefore more robust Cys-based native chemical ligation technique.The efficiency and capacity of this isoUb-based strategy is exemplified by the cost-effective synthesis of several linkage-and length-defined atypical Ub chains,including K27-linked tetra-Ub and K11/K48-branched tri-, tetra-, penta-, and hexa-Ubs.

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Modifying the Vicinity of the Isopeptide Bond To Reveal Differential Behavior of Ubiquitin Chains with Interacting Proteins: Organic Chemistry Applied to Synthetic Proteins

Cited by 27 publications

References 40 publications

Total Chemical Synthesis of ISGylated-Ubiquitin Hybrid Chain Assisted by Acetamidomethyl Derivatives with Dual Functions

Total Chemical Synthesis of ISGylated-Ubiquitin Hybrid Chain Assisted by Acetamidomethyl Derivatives with Dual Functions

A novel PEG-based solid support enables the synthesis of >50 amino-acid peptide thioesters and the total synthesis of a functional SUMO-1 peptide conjugate

Practical Chemical Synthesis of Atypical Ubiquitin Chains by Using an Isopeptide‐Linked Ub Isomer

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