scholarly journals Efficient protection and isolation of ubiquitylated proteins using tandem ubiquitin‐binding entities

EMBO Reports ◽  
2009 ◽  
Vol 10 (11) ◽  
pp. 1250-1258 ◽  
Author(s):  
Roland Hjerpe ◽  
Fabienne Aillet ◽  
Fernando Lopitz‐Otsoa ◽  
Valerie Lang ◽  
Patrick England ◽  
...  
Keyword(s):  
Ubiquitin Binding ◽  
Efficient Protection ◽  
Tandem Ubiquitin Binding Entities

Integrative analysis of the ubiquitin proteome isolated using Tandem Ubiquitin Binding Entities (TUBEs)

2012 ◽  
Vol 75 (10) ◽  
pp. 2998-3014 ◽  
Author(s):  
Fernando Lopitz-Otsoa ◽  
Eva Rodriguez-Suarez ◽  
Fabienne Aillet ◽  
Juan Casado-Vela ◽  
Valérie Lang ◽  
...  
Keyword(s):  
Integrative Analysis ◽  
Ubiquitin Binding ◽  
Tandem Ubiquitin Binding Entities

A Novel Luminescence-Based High-Throughput Approach for Cellular Resolution of Protein Ubiquitination Using Tandem Ubiquitin Binding Entities (TUBEs)

2020 ◽  
Vol 25 (4) ◽  
pp. 350-360
Author(s):  
Favour A. Akinjiyan ◽  
Aleem Fazal ◽  
Marc Hild ◽  
Rohan E. J. Beckwith ◽  
Nathan T. Ross ◽  
...  
Keyword(s):  
High Throughput ◽  
Ubiquitin Proteasome System ◽  
Live Cell ◽  
Novel Technologies ◽  
Substrate Protein ◽  
Protein Ubiquitination ◽  
Ubiquitin Binding ◽  
Ubiquitin Proteasome ◽  
Viable Approach ◽  
Tandem Ubiquitin Binding Entities

Protein turnover is highly regulated by the posttranslational process of ubiquitination. Deregulation of the ubiquitin proteasome system (UPS) has been implicated in cancer and neurodegenerative diseases, and modulating this system has proven to be a viable approach for therapeutic intervention. The development of novel technologies that enable high-throughput studies of substrate protein ubiquitination is key for UPS drug discovery. Conventional approaches for studying ubiquitination either have high protein requirements or rely on exogenous or modified ubiquitin moieties, thus limiting their utility. In order to circumvent these issues, we developed a high-throughput live-cell assay that combines the NanoBiT luminescence-based technology with tandem ubiquitin binding entities (TUBEs) to resolve substrate ubiquitination. To demonstrate the effectiveness and utility of this assay, we studied compound-induced ubiquitination of the G to S Phase Transition 1 (GSPT1) protein. Using this assay, we characterized compounds with varying levels of GSPT1 ubiquitination activity. This method provides a live-cell-based approach for assaying substrate ubiquitination that can be adapted to study the kinetics of ubiquitin transfer onto a substrate protein of interest. In addition, our results show that this approach is portable for studying the ubiquitination of target proteins with diverse functions.

Properties of natural and artificial proteins displaying multiple ubiquitin-binding domains

2010 ◽  
Vol 38 (1) ◽  
pp. 40-45 ◽  
Author(s):  
Fernando Lopitz-Otsoa ◽  
Manuel S. Rodríguez ◽  
Fabienne Aillet
Keyword(s):  
Effector Proteins ◽  
Target Protein ◽  
Polyubiquitin Chains ◽  
Artificial Proteins ◽  
Binding Domains ◽  
Cellular Factors ◽  
Evolutionary Advantage ◽  
Ubiquitin Binding ◽  
Tandem Ubiquitin Binding Entities

Ubiquitylation provides a rapid alternative to control the activity of crucial cellular factors through the remodelling of a target protein. Diverse ubiquitin chains are recognized by domains with affinity for UBDs (ubiquitin-binding domains) present in receptor/effector proteins. Interestingly, some proteins contain more than one UBD and the preservation of this structure in many species suggests an evolutionary advantage for this topology. Here, we review some typical proteins that naturally contain more than one UBD and emphasize how such structures contribute to the mechanism they mediate. Characteristics such as higher affinities for polyubiquitin chains and chain-linkage preferences can be replicated by the TUBEs (tandem ubiquitin-binding entities). Furthermore, TUBEs show two additional properties: protection of ubiquitylated substrates from deubiquitylating enzymes and interference with the action of the proteasome. Consequently, TUBEs behave as ‘ubiquitin traps’ that efficiently capture endogenous ubiquitylated proteins. Interpretations and hypothetical models proposed by different groups to understand the synchronous action of multiple UBDs are discussed herein.

Isolation of Ubiquitylated Proteins Using Tandem Ubiquitin-Binding Entities

2012 ◽  
pp. 173-183 ◽  
Author(s):  
Fabienne Aillet ◽  
Fernando Lopitz-Otsoa ◽  
Roland Hjerpe ◽  
Mónica Torres-Ramos ◽  
Valérie Lang ◽  
...  
Keyword(s):  
Ubiquitin Binding ◽  
Tandem Ubiquitin Binding Entities

Efficient protection of potato from diseases of various etiology under conditions of the Mosraw region

2019 ◽  
Vol 326 (3) ◽  
pp. 49-53
Author(s):  
M.A. Kuznetsova ◽  
◽  
A.N. Rogozhin ◽  
V.N. Demidova ◽  
T.I. Smetanina ◽  
...  
Keyword(s):  
Efficient Protection
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