Organometallic ruthenium anticancer complexes inhibit human peroxiredoxin I activity by binding to and inducing oxidation of its catalytic cysteine residue

Metallomics ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 546-555 ◽  
Author(s):  
Yu Lin ◽  
Jie Wang ◽  
Wei Zheng ◽  
Qun Luo ◽  
Kui Wu ◽  
...  
Keyword(s):  
Disulfide Bond ◽  
Cysteine Residue ◽  
Catalytic Cysteine ◽  
Anticancer Complexes ◽  
Ruthenium Arene ◽  
Peroxiredoxin I

Ruthenium arene anticancer complexes bind to human peroxiredoxin I, leading to oxidation of thiolate and failure of forming a disulfide bond.

scholarly journals Crystal structure of peroxiredoxin 3 from Vibrio vulnificus and its implications for scavenging peroxides and nitric oxide

IUCrJ ◽  
2018 ◽  
Vol 5 (1) ◽  
pp. 82-92 ◽  
Author(s):  
Jinsook Ahn ◽  
Kyung Ku Jang ◽  
Inseong Jo ◽  
Hasan Nurhasni ◽  
Jong Gyu Lim ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Nitric Oxide ◽  
Disulfide Bond ◽  
Vibrio Vulnificus ◽  
Cysteine Residue ◽  
Dimeric Interface ◽  
Catalytic Cysteine ◽  
New Type ◽  
Peroxidase Enzymes ◽  
Peroxiredoxin 3

Peroxiredoxins (Prxs) are ubiquitous cysteine-based peroxidase enzymes. Recently, a new type of Prx, VvPrx3, was identified in the pathogenic bacterium Vibrio vulnificus as being important for survival in macrophages. It employs only one catalytic cysteine residue to decompose peroxides. Here, crystal structures of VvPrx3 representing its reduced and oxidized states have been determined, together with an H2O2-bound structure, at high resolution. The crystal structure representing the reduced Prx3 showed a typical dimeric interface, called the A-type interface. However, VvPrx3 forms an oligomeric interface mediated by a disulfide bond between two catalytic cysteine residues from two adjacent dimers, which differs from the doughnut-like oligomers that appear in most Prxs. Subsequent biochemical studies showed that this disulfide bond was induced by treatment with nitric oxide (NO) as well as with peroxides. Consistently, NO treatment induced expression of the prx3 gene in V. vulnificus, and VvPrx3 was crucial for the survival of bacteria in the presence of NO. Taken together, the function and mechanism of VvPrx3 in scavenging peroxides and NO stress via oligomerization are proposed. These findings contribute to the understanding of the diverse functions of Prxs during pathogenic processes at the molecular level.

Ruthenium Arene Anticancer Complexes

2006 ◽  
pp. 39-64 ◽  
Author(s):  
Michael Melchart ◽  
Peter J. Sadler
Keyword(s):  
Anticancer Complexes ◽  
Ruthenium Arene

Transcription Inhibition by Organometallic Ruthenium - Arene Anticancer Complexes in Live Mammalian Cells

2012 ◽  
Vol 65 (9) ◽  
pp. 1271 ◽  
Author(s):  
Astrid Astarina ◽  
Mun Juinn Chow ◽  
Wee Han Ang
Keyword(s):  
Anticancer Agents ◽  
Mammalian Cells ◽  
Transcription Inhibition ◽  
Biological Targets ◽  
Arene Ligands ◽  
Arene Ligand ◽  
Anticancer Complexes ◽  
Covalent Adducts ◽  
The Spectator ◽  
Ruthenium Arene

Organometallic ruthenium–arene RAPTA complexes, currently being actively pursued as potential anticancer agents, interact with intracellular biological targets to form covalent adducts. Because their mode of action is still unclear, we investigated their binding with DNA and the ability of ruthenated-DNA adducts to elicit cellular responses such as transcription inhibition and repair. To investigate the influence of the spectator arene ligands on RAPTA activity, a novel RAPTA complex containing the bulky 1,3,5-triisopropylbenzene ligand was synthesized and characterized. Transcription experiments carried out in live mammalian cells using ruthenated plasmid probes revealed that increasing steric bulk of the arene ligand did not improve its ability to arrest transcription.

scholarly journals Activity of the Yap1 Transcription Factor in Saccharomyces cerevisiae Is Modulated by Methylglyoxal, a Metabolite Derived from Glycolysis

2004 ◽  
Vol 24 (19) ◽  
pp. 8753-8764 ◽  
Author(s):  
Kazuhiro Maeta ◽  
Shingo Izawa ◽  
Shoko Okazaki ◽  
Shusuke Kuge ◽  
Yoshiharu Inoue
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Transcription Factor ◽  
Disulfide Bond ◽  
Target Genes ◽  
Cysteine Residue ◽  
Cysteine Residues ◽  
Glyoxalase I ◽  
Toxic Metabolite ◽  
Basic Leucine Zipper

ABSTRACT Methylglyoxal (MG) is synthesized during glycolysis, although it inhibits cell growth in all types of organisms. Hence, it has long been asked why such a toxic metabolite is synthesized in vivo. Glyoxalase I is a major enzyme detoxifying MG. Here we show that the Yap1 transcription factor, which is critical for the oxidative-stress response in Saccharomyces cerevisiae, is constitutively concentrated in the nucleus and activates the expression of its target genes in a glyoxalase I-deficient mutant. Yap1 contains six cysteine residues in two cysteine-rich domains (CRDs), i.e., three cysteine residues clustering near the N terminus (n-CRD) and the remaining three cysteine residues near the C terminus (c-CRD). We reveal that any of the three cysteine residues in the c-CRD is sufficient for MG to allow Yap1 to translocate into the nucleus and to activate the expression of its target gene. A Yap1 mutant possessing only one cysteine residue in the c-CRD but no cysteine in the n-CRD and deletion of the basic leucine zipper domain can concentrate in the nucleus with MG treatment. However, substitution of all the cysteine residues in Yap1 abolishes the ability of this transcription factor to concentrate in the nucleus following MG treatment. The redox status of Yap1 is substantially unchanged, and protein(s) interaction with Yap1 through disulfide bond is hardly detected in cells treated with MG. Collectively, neither intermolecular nor intramolecular disulfide bond formation seems to be involved in Yap1 activation by MG. Moreover, we show that nucleocytoplasmic localization of Yap1 closely correlates with growth phase and intracellular MG level. We propose a novel regulatory pathway underlying Yap1 activation by a natural metabolite in the cell.

scholarly journals In VivoFormation of the Protein Disulfide Bond That Enhances the Thermostability of Diphosphomevalonate Decarboxylase, an Intracellular Enzyme from the Hyperthermophilic Archaeon Sulfolobus solfataricus

2015 ◽  
Vol 197 (21) ◽  
pp. 3463-3471 ◽  
Author(s):  
Ai Hattori ◽  
Hideaki Unno ◽  
Shuichiro Goda ◽  
Kento Motoyama ◽  
Tohru Yoshimura ◽  
...  
Keyword(s):  
Disulfide Bond ◽  
Mevalonate Pathway ◽  
Sulfolobus Solfataricus ◽  
Cysteine Residue ◽  
Remarkable Feature ◽  
Intracellular Enzyme ◽  
Content Type ◽  
Hyperthermophilic Archaeon ◽  
Whole Cells ◽  
Physiological Importance

ABSTRACTIn the present study, the crystal structure of recombinant diphosphomevalonate decarboxylase from the hyperthermophilic archaeonSulfolobus solfataricuswas solved as the first example of an archaeal and thermophile-derived diphosphomevalonate decarboxylase. The enzyme forms a homodimer, as expected for most eukaryotic and bacterial orthologs. Interestingly, the subunits of the homodimer are connected via an intersubunit disulfide bond, which presumably formed during the purification process of the recombinant enzyme expressed inEscherichia coli. When mutagenesis replaced the disulfide-forming cysteine residue with serine, however, the thermostability of the enzyme was significantly lowered. In the presence of β-mercaptoethanol at a concentration where the disulfide bond was completely reduced, the wild-type enzyme was less stable to heat. Moreover, Western blot analysis combined with nonreducing SDS-PAGE of the whole cells ofS. solfataricusproved that the disulfide bond was predominantly formed in the cells. These results suggest that the disulfide bond is required for the cytosolic enzyme to acquire further thermostability and to exert activity at the growth temperature ofS. solfataricus.IMPORTANCEThis study is the first report to describe the crystal structures of archaeal diphosphomevalonate decarboxylase, an enzyme involved in the classical mevalonate pathway. A stability-conferring intersubunit disulfide bond is a remarkable feature that is not found in eukaryotic and bacterial orthologs. The evidence that the disulfide bond also is formed inS. solfataricuscells suggests its physiological importance.

scholarly journals Monothiol and dithiol glutaredoxin-1 from Clostridium oremlandii: identification of domain-swapped structures by NMR, X-ray crystallography and HDX mass spectrometry

IUCrJ ◽  
2020 ◽  
Vol 7 (6) ◽  
pp. 1019-1027
Author(s):  
Kitaik Lee ◽  
Kwon Joo Yeo ◽  
Sae Hae Choi ◽  
Eun Hye Lee ◽  
Bo Keun Kim ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Disulfide Bond ◽  
Protein Function ◽  
Single Point ◽  
Cysteine Residue ◽  
Single Point Mutation ◽  
Protein Oligomerization ◽  
X Ray ◽  
X Ray Crystallography ◽  
Hydrogen Deuterium

Protein dimerization or oligomerization resulting from swapping part of the protein between neighboring polypeptide chains is known to play a key role in the regulation of protein function and in the formation of protein aggregates. Glutaredoxin-1 from Clostridium oremlandii (cGrx1) was used as a model to explore the formation of multiple domain-swapped conformations, which were made possible by modulating several hinge-loop residues that can form a pivot for domain swapping. Specifically, two alternative domain-swapped structures were generated and analyzed using nuclear magnetic resonance (NMR), X-ray crystallography, circular-dichroism spectroscopy and hydrogen/deuterium-exchange (HDX) mass spectrometry. The first domain-swapped structure (β3-swap) was formed by the hexameric cGrx1–cMsrA complex. The second domain-swapped structure (β1-swap) was formed by monothiol cGrx1 (C16S) alone. In summary, the first domain-swapped structure of an oxidoreductase in a hetero-oligomeric complex is presented. In particular, a single point mutation of a key cysteine residue to serine led to the formation of an intramolecular disulfide bond, as opposed to an intermolecular disulfide bond, and resulted in modulation of the underlying free-energy landscape of protein oligomerization.

scholarly journals Itaconate is a covalent inhibitor of the Mycobacterium tuberculosis isocitrate lyase

2020 ◽  
Author(s):  
Brooke X. C. Kwai ◽  
Annabelle J. Collins ◽  
Martin J. Middleditch ◽  
Jonathan Sperry ◽  
Ghader Bashiri ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Catalytic Mechanism ◽  
Isocitrate Lyase ◽  
Cysteine Residue ◽  
Covalent Adduct ◽  
Catalytic Cysteine ◽  
Covalent Inhibitor

Mycobacterium tuberculosis isocitrate lyases (ICLs) form a covalent adduct with itaconate through their catalytic cysteine residue. These results reveal atomic details of itaconate inhibition and provide insights into the catalytic mechanism of ICLs.

Organometallic Chemistry, Biology and Medicine: Ruthenium Arene Anticancer Complexes

ChemInform ◽  
2006 ◽  
Vol 37 (4) ◽  
Author(s):  
Yaw Kai Yan ◽  
Michael Melchart ◽  
Abraha Habtemariam ◽  
Peter J. Sadler
Keyword(s):  
Organometallic Chemistry ◽  
Anticancer Complexes ◽  
Ruthenium Arene

scholarly journals The rhodanese RhdA helps Azotobacter vinelandii in maintaining cellular redox balance

2010 ◽  
Vol 391 (7) ◽  
Author(s):  
William Remelli ◽  
Angelo Cereda ◽  
Jutta Papenbrock ◽  
Fabio Forlani ◽  
Silvia Pagani
Keyword(s):  
Oxidative Stress ◽  
Stress Condition ◽  
Azotobacter Vinelandii ◽  
Redox Balance ◽  
Cysteine Residue ◽  
Structural Peculiarity ◽  
Cellular Redox ◽  
Catalytic Cysteine ◽  
Transcript Profiles

AbstractThe tandem domain rhodanese-homology protein RhdA ofAzotobacter vinelandiishows an active-site loop structure that confers structural peculiarity in the environment of its catalytic cysteine residue. Thein vivoeffects of the lack of RhdA were investigated using anA. vinelandiimutant strain (MV474) in which therhdAgene was disrupted by deletion. Here, by combining analytical measurements and transcript profiles, we show that deletion of therhdAgene generates an oxidative stress condition to whichA. vinelandiiresponds by activating defensive mechanisms. In conditions of growth in the presence of the superoxide generator phenazine methosulfate, a stressor-dependent induction ofrhdAgene expression was observed, thus highlighting that RhdA is important forA. vinelandiito sustain oxidative stress. The potential of RhdA to buffer general levels of oxidants inA. vinelandiicells via redox reactions involving its cysteine thiol is discussed.

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