One-shot NMR analysis of microbial secretions identifies highly potent proteasome inhibitor

Stein, Manuel; Beck, Philipp; Kaiser, Markus; Dudler, Robert; Becker, Christian F. W.; Groll, M.

doi:10.1073/pnas.1211423109

Cited by 59 publications

(62 citation statements)

References 32 publications

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“…Furthermore, GlbA as well as a variant named cepafungin I, the strongest proteasome inhibitor known to date, were isolated from Ph. luminescens cultures [67]. Similarly, GlbA and derivatives named luminmycins have been isolated from Ph.…”

Section: Whereas Disrupted Nonfunctional Glb-like Gene Clusters Are Pmentioning

confidence: 99%

The role of bacterial phytotoxins in inhibiting the eukaryotic proteasome

Dudler

2014

Trends in Microbiology

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The ubiquitin-26S proteasome degradation system (UPS) plays a pivotal role in almost all aspects of plant life, including defending against pathogens. Although the proteasome is important for plant immunity, it has been found to be also exploited by pathogens using effectors to increase their virulence. Recent work on the XopJ effector and syringolin A/syrbactins has highlighted host proteasome inhibition as a virulence strategy of pathogens. This review will focus on these recent developments. AbstractThe ubiquitin-26S proteasome degradation system (UPS) plays a pivotal role in almost all aspects of plant life, including defending against pathogens. While the proteasome is important for plant immunity, it has been found to also be exploited by pathogens using effectors to increase their virulence. Recent work on the XopJ effector and syringolin A/syrbactins has highlighted host proteasome inhibition as a virulence strategy of pathogens. This review will focus on these recent developments. The ubiquitin-26S proteasome degradation system (UPS) and plant immunityBesides the lysosome, the UPS is the main protein degradation system of eukaryotic cells that not only destructs misfolded proteins but is also involved in many cellular processes by degrading proteins regulating such processes [1,2]. Proteins destined for destruction by the proteasome become polyubiquitinated by a reaction cascade involving three enzymes designated E1, E2, and E3. Ubiquitin is activated in an ATP-dependent way by E1 and transferred to E2, from which it is normally directly transferred to a lysine residue of the target protein by E3, which, by selecting target proteins, confers specificity to the UPS [3].Polyubiquitination is achieved by multiple rounds in which further ubiquitin units are conjugated to an internal lysine residue of ubiquitin. Polyubiquitinated proteins are substrates for the proteasome, which is composed of two 19S regulatory particles (RP) capping a 20S core particle (CP). The RPs recognize polyubiquitinated proteins, which become deubiquitinated and unfolded in an ATP-dependent reaction, and regulate access of unfolded substrates to the proteolytic channel of the CP. The channel is formed by four stacked sevenmembered rings. The two identical outer rings consist of seven different α-subunits, whereas 3 the two identical inner rings consist of seven different β-subunits, three of which have proteolytic activities: the β1 subunit exhibits a caspase-like activity (CL), cleaving protein substrates after acidic residues, while the β2 and β5 subunits have trypsin-like (TL) and chymotrypsin-like (ChTL) activities cleaving after basic and hydrophobic residues, respectively. In all three catalytic subunits, the active site residue is the N-terminal threonineThe UPS is involved in the signaling pathways of nearly all plant hormones, including salicylic acid (SA), jasmonic acid (JA), and ethylene (ET), hormones of particular importance for plant defense against pathogens [5]. SA plays a prominent role in defense reactio...

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Section: Whereas Disrupted Nonfunctional Glb-like Gene Clusters Are Pmentioning

confidence: 99%

The role of bacterial phytotoxins in inhibiting the eukaryotic proteasome

Dudler

2014

Trends in Microbiology

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“…The MS 2 spectrum indicates that azidation of glidobactins only took place on the reactive site that is also targeted by the proteasome (Supporting Information File 1, Figure S11) [24]. …”

Section: Resultsmentioning

confidence: 99%

Solid-phase enrichment and analysis of electrophilic natural products

Wesche¹,

He²,

Bode³

2017

Beilstein J. Org. Chem.

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In search for new natural products, which may lead to the development of new drugs for all kind of applications, novel methods are needed. Here we describe the identification of electrophilic natural products in crude extracts via their reactivity against azide as a nucleophile followed by their subsequent enrichment using a cleavable azide-reactive resin (CARR). Using this approach, natural products carrying epoxides and α,β-unsaturated enones as well as several unknown compounds were identified in crude extracts from entomopathogenic Photorhabdus bacteria.

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“…One such reporter used small peptides that, when degraded by the proteasome, would exhibit a decreased FRET-behavior and direct measurement of proteasome activity (31). Finally, a novel reporting technique incorporated a short peptide along with nuclear magnetic resonance (NMR) to detect cleavage of an isotopically labeled reporter (32). This enabled detection of proteasome activity even in optically dense environments, and has been used to discover potential proteasome inhibitors in the secretions of Photorhabdus luminescens .…”

Section: Measuring Proteasome Activitymentioning

confidence: 99%

Measuring Activity in the Ubiquitin–Proteasome System: From Large Scale Discoveries to Single Cells Analysis

Melvin

Woss

Park

et al. 2013

Cell Biochem Biophys

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The ubiquitin proteasome system (UPS) is the primary pathway responsible for the recognition and degradation of misfolded, damaged, or tightly regulated proteins in addition to performing essential roles in DNA repair, cell cycle regulation, cell migration, and the immune response. While traditional biochemical techniques have proven useful in the identification of key proteins involved in this pathway, the implementation of novel reporters responsible for measuring enzymatic activity of the UPS have provided valuable insight into the effectiveness of therapeutics and role of the UPS in various human diseases such as multiple myeloma and Huntington’s disease. These reporters, usually consisting of a recognition sequences fused to an analytical handle, are designed to specifically evaluate enzymatic activity of certain members of the UPS including the proteasome, E3 ubiquitin ligases, and deubiquitinating enzymes (DUBs). This review highlights the more commonly used reporters employed in a variety of scenarios ranging from high-throughput screening of novel inhibitors to single cell microscopy techniques measuring E3 ligase or proteasome activity. Finally, recent work is presented highlighting the development of novel degron-based substrate designed to overcome the limitations of current reporting techniques in measuring E3 ligase and proteasome activity in patient samples.

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One-shot NMR analysis of microbial secretions identifies highly potent proteasome inhibitor

Cited by 59 publications

References 32 publications

The role of bacterial phytotoxins in inhibiting the eukaryotic proteasome

The role of bacterial phytotoxins in inhibiting the eukaryotic proteasome

Solid-phase enrichment and analysis of electrophilic natural products

Measuring Activity in the Ubiquitin–Proteasome System: From Large Scale Discoveries to Single Cells Analysis

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