Functional characterization of a SUMO deconjugating protease of Plasmodium falciparum using newly identified small molecule inhibitors.

Published on Jun 24, 2011in Chemistry & Biology
· DOI :10.1016/J.CHEMBIOL.2011.04.010
Elizabeth L. Ponder8
Estimated H-index: 8
(Stanford University),
Victoria E. Albrow13
Estimated H-index: 13
(Stanford University)
+ 11 AuthorsMatthew Bogyo88
Estimated H-index: 88
(Stanford University)
Sources
Abstract
Summary Small ubiquitin-related modifier (SUMO) is implicated in the regulation of numerous biological processes including transcription, protein localization, and cell cycle control. Protein modification by SUMO is found in Plasmodium falciparum ; however, its role in the regulation of the parasite life cycle is poorly understood. Here we describe functional studies of a SUMO-specific protease (SENP) of P. falciparum , PfSENP1 (PFL1635w). Expression of the catalytic domain of PfSENP1 and biochemical profiling using a positional scanning substrate library demonstrated that this protease has unique cleavage sequence preference relative to the human SENPs. In addition, we describe a class of small molecule inhibitors of this protease. The most potent lead compound inhibited both recombinant PfSENP1 activity and P. falciparum replication in infected human blood. These studies provide valuable new tools for the study of SUMOylation in P. falciparum .
📖 Papers frequently viewed together
1,376 Citations
50 Citations
393 Citations
References36
Newest
#1Victoria E. Albrow (Stanford University)H-Index: 13
#2Elizabeth L. Ponder (Stanford University)H-Index: 8
Last. Matthew Bogyo (Stanford University)H-Index: 88
view all 7 authors...
Summary Sentrin specific proteases (SENPs) are responsible for activating and deconjugating SUMO (Small Ubiquitin-like MOdifier) from target proteins. It remains difficult to study this posttranslational modification due to the lack of reagents that can be used to block the removal of SUMO from substrates. Here, we describe the identification of small molecule SENP inhibitors and active site probes containing aza-epoxide and acyloxymethyl ketone (AOMK) reactive groups. Both classes of compounds ...
50 CitationsSource
#1Heather J. Painter (Princeton University)H-Index: 17
#2Tracey L. Campbell (Princeton University)H-Index: 3
Last. Manuel Llinás (Princeton University)H-Index: 49
view all 3 authors...
Malaria is caused by protozoan parasites of the genus Plasmodium and involves infection of multiple hosts and cell types during the course of an infection. To complete its complex life cycle the parasite requires strict control of gene regulation for survival and successful propagation. Thus far, the Apicomplexan AP2 (ApiAP2) family of DNA-binding proteins is the sole family of proteins to have surfaced as candidate transcription factors in all apicomplexan species. Work from several laboratorie...
153 CitationsSource
#1Jake Baum (WEHI: Walter and Eliza Hall Institute of Medical Research)H-Index: 40
#2Anthony T. Papenfuss (WEHI: Walter and Eliza Hall Institute of Medical Research)H-Index: 53
Last. Tania F. de Koning-Ward (WEHI: Walter and Eliza Hall Institute of Medical Research)H-Index: 32
view all 9 authors...
Techniques for targeted genetic disruption in Plasmodium, the causative agent of malaria, are currently intractable for those genes that are essential for blood stage development. The ability to use RNA interference (RNAi) to silence gene expression would provide a powerful means to gain valuable insight into the pathogenic blood stages but its functionality in Plasmodium remains controversial. Here we have used various RNA-based gene silencing approaches to test the utility of RNAi in malaria p...
162 CitationsSource
#1Mary B. KroetzH-Index: 5
#2Dan Su (Yale University)H-Index: 4
Last. Mark Hochstrasser (Yale University)H-Index: 78
view all 3 authors...
The SUMO protein is covalently attached to many different substrates throughout the cell. This modification is rapidly reversed by SUMO proteases. The Saccharomyces cerevisiae SUMO protease Ulp2 is a nuclear protein required for chromosome stability and cell cycle restart after checkpoint arrest. Ulp2 is related to the human SENP6 protease, also a nuclear protein. All members of the Ulp2/SENP6 family of SUMO proteases have large but poorly conserved N-terminal domains (NTDs) adjacent to the cata...
29 CitationsSource
#1Helle D. Ulrich (London Research Institute)H-Index: 30
Modifications of the eukaryotic sliding clamp, proliferating cell nuclear antigen (PCNA), by ubiquitin and the ubiquitin-related protein SUMO, are well known to influence the choice of pathways for the processing of DNA lesions during replication. Over the past few years, significant progress has been made not only with respect to the molecular consequences that each of the modifications has for the properties of PCNA, but also in terms of the cellular signals that elicit the ubiquitylation or s...
140 CitationsSource
#1Edward T.H. Yeh (University of Texas MD Anderson Cancer Center)H-Index: 93
The small ubiquitin-like modifier (SUMO) is a ubiquitin-like protein that covalently modifies a large number of cellular proteins. SUMO modification has emerged as an important regulatory mechanism for protein function and localization. SUMOylation is a dynamic process that is mediated by activating (E1), conjugating (E2), and ligating (E3) enzymes and readily reversed by a family of ubiquitin-like protein-specific proteases (Ulp) in yeast and sentrin/SUMO-specific proteases (SENP) in human. Thi...
362 CitationsSource
#1Laurence Braun (UJF: Joseph Fourier University)H-Index: 15
#2Dominique Cannella (UJF: Joseph Fourier University)H-Index: 14
Last. Mohamed-Ali Hakimi (UJF: Joseph Fourier University)H-Index: 39
view all 7 authors...
SUMOylation, the reversible covalent attachment of small ubiquitin-like modifier (SUMO) peptides has emerged as an important regulator of target protein function. Here we show, by characterization of the Toxoplasma gondii SUMO pathway, that the SUMO conjugation system operates in apicomplexan parasites. A gene encoding the SUMO tag was discovered as were genes encoding the various enzymes required for SUMO processing, ligation and release. Various SUMO conjugates were immuno-detected and by mean...
43 CitationsSource
#1Bernardo J. Foth (NTU: Nanyang Technological University)H-Index: 19
#2Neng Zhang (NTU: Nanyang Technological University)H-Index: 3
Last. Zbynek Bozdech (NTU: Nanyang Technological University)H-Index: 38
view all 5 authors...
Background: Malaria is a one of the most important infectious diseases and is caused by parasitic protozoa of the genus Plasmodium. Previously, quantitative characterization of the P. falciparum transcriptome demonstrated that the strictly controlled progression of these parasites through their intra-erythrocytic developmental cycle is accompanied by a continuous cascade of gene expression. Although such analyses have proven immensely useful, the correlations between abundance of transcripts and...
100 CitationsSource
Small ubiquitin-like modifier (SUMO) proteases regulate the abundance and lifetime of SUMO-conjugated substrates by antagonizing reactions catalyzed by SUMO-conjugating enzymes. Six SUMO proteases constitute the human SENP/ULP protease family (SENP1-3 and SENP5-7). SENP6 and SENP7 include the most divergent class of SUMO proteases, which also includes the yeast enzyme ULP2. We present the crystal structure of the SENP7 catalytic domain at a resolution of 2.4A. Comparison with structures of human...
93 CitationsSource
#1Marcin Drag (Wrocław University of Technology)H-Index: 37
#2Jowita Mikolajczyk (Sanford-Burnham Institute for Medical Research)H-Index: 13
Last. Guy S. Salvesen (Sanford-Burnham Institute for Medical Research)H-Index: 127
view all 7 authors...
DUBs (deubiquitinating enzymes) are a family of proteases responsible for the specific removal of ubiquitin attached to target proteins and thus control the free cellular pools of this molecule. DUB activity is usually assayed using full-length ubiquitin, and these enzymes generally show low activity towards small substrates that constitute the P4–P1 LRGG (Lys-Arg-Gly-Gly) C-terminal motif of ubiquitin. To gain insight into the C-terminal recognition region of ubiquitin by DUBs, we synthesized p...
49 CitationsSource
Cited By35
Newest
#1Antti Kukkula (TYKS: Turku University Hospital)
#2Veera K. Ojala (TYKS: Turku University Hospital)
Last. Maria Sundvall (TYKS: Turku University Hospital)H-Index: 14
view all 6 authors...
SUMOylation is a dynamic and reversible post-translational modification, characterized more than 20 years ago, that regulates protein function at multiple levels. Key oncoproteins and tumor suppressors are SUMO substrates. In addition to alterations in SUMO pathway activity due to conditions typically present in cancer, such as hypoxia, the SUMO machinery components are deregulated at the genomic level in cancer. The delicate balance between SUMOylation and deSUMOylation is regulated by SENP enz...
Source
#1R.K. Mishra (Indian Institutes of Science Education and Research)
#2Jai Shankar Singh (IITB: Indian Institute of Technology Bombay)H-Index: 2
Last. Ashutosh Kumar (IITB: Indian Institute of Technology Bombay)H-Index: 5
view all 8 authors...
Abstract null The endoparasitic pathogen, Plasmodium falciparum (Pf), modulates protein-protein interactions to employ post-translational modifications like SUMOylation in order to establish successful infections. The interaction between E1 and E2 (Ubc9) enzymes governs species specificity in the Plasmodium SUMOylation pathway. Here, we demonstrate that a unidirectional cross-species interaction exists between Pf-SUMO and Human-E2, whereas Hs-SUMO1 failed to interact with Pf-E2. Biochemical and ...
Source
#1Majid Dousti (Shiraz University of Medical Sciences)H-Index: 4
#2Raúl Manzano-Román (CSIC: Spanish National Research Council)H-Index: 22
Last. Gholamreza Hatam (Shiraz University of Medical Sciences)H-Index: 25
view all 7 authors...
There is no effective vaccine against malaria; therefore, chemotherapy is to date only choice to fight against this infectious disease. However, there are growing evidences of drug-resistance mechanisms in malaria treatments. Therefore, the identification of new drug targets is an urgent need for the clinic management of the disease. Proteomic approaches offer the chance of determining the effects of antimalarial drugs on the proteome of Plasmodium parasites. Accordingly, we here review the effe...
1 CitationsSource
Source
Post-translational protein regulation allows for fine-tuning of cellular functions and involves a wide range of modifications, including ubiquitin and ubiquitin-like modifiers (Ubls). The dynamic balance of Ubl conjugation and removal shapes the fates of target substrates, in turn modulating various cellular processes. The mechanistic aspects of Ubl pathways and their biological roles have been largely established in yeast, plants, and mammalian cells. However, these modifiers may be utilised di...
5 CitationsSource
#1Urs Lindenmann (Zurich University of Applied Sciences/ZHAW)H-Index: 2
#2Michael Brand (Zurich University of Applied Sciences/ZHAW)H-Index: 2
Last. Rainer Riedl (Zurich University of Applied Sciences/ZHAW)H-Index: 13
view all 8 authors...
Sentrin-specific proteases (SENPs) are responsible for the maturation of small ubiquitin-like modifiers (SUMOs) and the deconjugation of SUMOs from their substrate proteins. Studies on prostate cancer revealed an overexpression of SENP1, which promotes prostate cancer progression as well as metastasis. Therefore, SENP1 has been identified as a novel drug target against prostate cancer. Herein, we report the discovery and biological evaluation of potent and selective SENP1 inhibitors. A structure...
3 CitationsSource
#1Natasha Stella Tibon (International Medical University)H-Index: 1
#2Chew Hee Ng (International Medical University)H-Index: 14
Last. Siew Lee Cheong (International Medical University)H-Index: 3
view all 3 authors...
Abstract Discovery and development of antimalarial drugs have long been dominated by single-target therapy. Continuous effort has been made to explore and identify different targets in malaria parasite crucial for the malaria treatment. The single-target drug therapy was initially successful, but it was later supplanted by combination therapy with multiple drugs to overcome drug resistance. Emergence of resistant strains even against the combination therapy has warranted a review of current anti...
14 CitationsSource
#1Shiyu Chen (Stanford University)H-Index: 12
#2Joshua J. Yim (Stanford University)H-Index: 13
Last. Matthew Bogyo (Stanford University)H-Index: 88
view all 3 authors...
Proteases are regulators of diverse biological pathways including protein catabolism, antigen processing and inflammation, as well as various disease conditions, such as malignant metastasis, viral infection and parasite invasion. The identification of substrates of a given protease is essential to understand its function and this information can also aid in the design of specific inhibitors and active site probes. However, the diversity of putative protein and peptide substrates makes connectin...
8 CitationsSource
#1Edgar Deu (Francis Crick Institute)H-Index: 16
Malaria is a devastating parasitic disease affecting half of the world's population. The rapid emergence of resistance against new antimalarial drugs, including artemisinin-based therapies, has made the development of drugs with novel mechanisms of action extremely urgent. Proteases are enzymes proven to be well suited for target-based drug development due to our knowledge of their enzymatic mechanisms and active site structures. More importantly, Plasmodium proteases have been shown to be invol...
41 CitationsSource
Experimental immunization with radiation attenuated sporozoites (RAS) and genetically attenuated sporozoites has proved to be a promising approach for malaria vaccine development. However, parasite biomarkers of growth attenuation and enhanced immune protection in response to radiation remain poorly understood. Here, we report on the effect of an attenuating dose of γ-irradiation (15 krad) on the Plasmodium falciparum sporozoite (PfSPZ) ultrastructure by electron microscopy, growth rate of liver...
11 CitationsSource