A strategy for modulation of enzymes in the ubiquitin system.
Andreas Ernst,George V. Avvakumov,Jiefei Tong,Yihui Fan,Yanling Zhao,Philipp Alberts,Avinash Persaud,John R. Walker,Ana-Mirela Neculai,Dante Neculai,Andrew Vorobyov,Pankaj Garg,Linda G. Beatty,Pak-Kei Chan,Yu Chi Juang,Marie-Claude Landry,Christina Yeh,Christina Yeh,Elton Zeqiraj,Konstantina Karamboulas,Abdellah Allali-Hassani,Masoud Vedadi,Mike Tyers,Mike Tyers,Jason Moffat,Frank Sicheri,Frank Sicheri,Laurence Pelletier,Laurence Pelletier,Daniel Durocher,Daniel Durocher,Brian Raught,Daniela Rotin,Jianhua Yang,Michael Moran,Sirano Dhe-Paganon,Sirano Dhe-Paganon,Sachdev S. Sidhu +37 more
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TLDR
This work used massively diverse combinatorial libraries of ubiquitin variants to develop inhibitors of four deubiquitinases (DUBs) and analyzed the DUB-inhibitor complexes with crystallography to report a method to target the myriad enzymes that govern ubiquitination of protein substrates.Abstract:
The ubiquitin system regulates virtually all aspects of cellular function. We report a method to target the myriad enzymes that govern ubiquitination of protein substrates. We used massively diverse combinatorial libraries of ubiquitin variants to develop inhibitors of four deubiquitinases (DUBs) and analyzed the DUB-inhibitor complexes with crystallography. We extended the selection strategy to the ubiquitin conjugating (E2) and ubiquitin ligase (E3) enzymes and found that ubiquitin variants can also enhance enzyme activity. Last, we showed that ubiquitin variants can bind selectively to ubiquitin-binding domains. Ubiquitin variants exhibit selective function in cells and thus enable orthogonal modulation of specific enzymatic steps in the ubiquitin system.read more
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On-Demand Detachment of Succinimides on Cysteine to Facilitate (Semi)Synthesis of Challenging Proteins.
Ganga B. Vamisetti,Gandhesiri Satish,Prasad Sulkshane,Guy Mann,Michael H. Glickman,Ashraf Brik +5 more
TL;DR: The advantages of the newly developed method for the protection and deprotection of native cysteine with a succinimide group in a peptide fragment derived from thioredoxin-1 obtained via intein based expression to enable ligation/desulfurization and subsequent disulfide bond formation in a one-pot process.
Journal ArticleDOI
Peptide Level Immunoaffinity Enrichment Enhances Ubiquitination Site Identification on Individual Proteins
Veronica G. Anania,Victoria Pham,XiaoDong Huang,Alexandre Masselot,Jennie R. Lill,Donald S. Kirkpatrick +5 more
TL;DR: The utility of immunoaffinity peptide enrichment for single protein ubiquitination site analysis is investigated and insights into the ubiquitinations of HER2, DVL2, and proteins in the T-cell receptor complex are provided.
Journal ArticleDOI
Probing ubiquitin and SUMO conjugation and deconjugation
Huib Ovaa,Alfred C.O. Vertegaal +1 more
TL;DR: The current state of the art in this field is discussed and what is needed to unravel the connections between different components of the conjugation machineries for ubiquitylation and SUMOylation is pointed out.
Journal ArticleDOI
Allosteric Modulation of Binding Specificity by Alternative Packing of Protein Cores
Moshe Ben-David,Haiming Huang,Mark G. F. Sun,Carles Corbi-Verge,Evangelia Petsalaki,Ke Liu,David Gfeller,Pankaj Garg,Wolfram Tempel,Irina Sochirca,Julia M. Shifman,Alan R. Davidson,Jinrong Min,Philip M. Kim,Sachdev S. Sidhu +14 more
TL;DR: The results show that alternative core packing and consequent allosteric modulation of binding interfaces could be used to engineer proteins with novel functions.
Journal ArticleDOI
Modes of allosteric regulation of the ubiquitination machinery.
TL;DR: Recent examples of ubiquitin itself as an allosteric effector of RING and RBR E3 ligases, as well as advances in the understanding of allosterics regulatory elements within HECT E3ligases are considered.
References
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