A warm welcome from the Saarland Bioorganic Chemistry, School of Pharmacy Saarland University, Saarbrücken, Germany Claus Jacob intracellular signalling via the cellular thiolstat From antioxidant activity to complex redox modulation and 3 Organochalcogen Compounds in Chemistry and Biology 1 • What’s next ?? • Intracellular diagnostics • From nutrition to wider applications • Interferences using sulfur-containing nutrachemicals • Redox control and the cellular thiolstat Overview of today’s talk • 80% Diallyldisulfide is not 100% Diallyldisulfide • Polysulfides are not polysulfanes • Redox is not a Red Ox • Johanna Wanka is not Willy Wonka • Saarland is not Saharaland • Ischia is not Ischias Nomen est omen 4 2 7 and tellurium based molecules • Many ROS react effectively, yet also selectively with sulfur, selenium • Exchange reactions are common with oxygen, sulfur and selenium oxidation states (e.g. exchange reactions) transfer and nucleophilic substitutions with simultaneous changes of • Other mechanisms include radical reactions, atom transfer, hydride but not necessarily always preferred • In Biology, electron transfer is common (molecular) mechanism • Redox processes follow a certain The Special Relationship thiolstat Redox control and the cellular 5 +2 (N) -1 (O) NOS O2x- + xNO UV-radiation, causes skin cancer Inflammation, metabolism Inflammation, metabolism Fenton reaction Occurrence cysteine redox transformations lead to 8 6 biochemical Sensing, Signalling cascades and feedback loops are possible Most cysteine redox transformations are reversible consequences Most Sulfur species may react by a variety of (redox) mechanisms Up to 10 oxidation states of sulfur known in vivo High abundance of sulfur in vivo (often millimolar concentrations) Sulfur in (human) Biology ONOO- xNO -1 HOx Hydroxyl radical Nitric oxide Peroxynitrite -1 H2 O2 Peroxide -0.5 O2x- Superoxide 0 1O 2 Ox. State Singlet Oxygen Reactive Species Reactive Species and Oxidative Stress Jacob et al., ChemBioChem 2011 Trapping and hunting sulfur modifications Giles et al., Free Radic. Biol. Med. 2001; Giles et al., Biochem. Biophys. Res. Commun. 2003, Chem. Biol. 2003; Jacob and Sies et al., Angew. Chem. 2003 Reactive Sulfur Species in the human body 11 9 Jamier et al., Chem. Eur. J. 2010 Schafer and Buettner Free Radical Biol. Med. 2001 Once upon a time: Electrochemical potential, GSSG:GSH ratio and cellular responses • Metal-sulfur interactions (metal-redox interdependence) • “Odd” modifications (perthiols, trisulfanes) 12 • Over-oxidation to form the cellular ‘sulfenome’ (sulfinic, sulfonic acids) • S-nitrosation (RSNO formation and •NO release) interactions) and - as reverse - erratic cleavage • Disulfide bond formation (including reversible protein-protein • S-thiolation of cysteine residues (mostly S-glutathiolation) • Thiol oxidation Posttranslational modifications in peptides, proteins and enzymes 10 (especially cysteinebased redox proteins involved in cellular signalling) And many more..... Fructose-1,6-biphosphatase Glucose-6-phosphate dehydrogenase Necrosis Widespread damage and inactivation of proteins/enzymes PrSSG PrSSPr Cellular Thiolstat Decrease of ROS level Antioxidant feedback Widespread S-thiolation, oxidation and overoxidation Increase of ROS levels PrSSG PrSSPr Apoptosis Activation of proapoptotic pathways Cellular signaling Protein/enzyme regulation Shift of intracellular redox state to more oxidizing potentials S-thiolation / oxidation of cysteine-containing proteins/enzymes Activation of antioxidant response pathways Jacob 2011, Jacob et al. 2011 Signalling and Control via the Cellular Thiolstat 15 after Ghezzi et al., Redox Signaling and Regulation in Biology and Medicine, Wiley VCH 2009 3-hydroxy-3-methylglutaryl-CoA reductase Glycogen synthase I Phosphorylase phosphatase Thioredoxin β-Ketoacylthiolase Guanylate cyclase Pyruvate dehydrogenase Phosphofructokinase Collagenase Trypsin Hexokinase Activated by protein thiolation Pyruvate kinase 13 Inactivated by protein thiolation Proteins and enzymes affected by Sthiolation/dethiolation 14 Interferences using sulfur-containing nutrachemicals Is it possible to interfere with the cellular thiolstat ?? and regulation. 16 responses – in one word: effective cellular signalling appropriate, measured and largely reversible cellular redox manipulation of cysteine residues enables fast, Modulation of protein function and enzyme activity by “Cellular Thiolstat” From the “Sulfur Redoxome” towards the SH S S S NH2 S S O S COOH NH2 COOH S S S S S S S S SH S O S S S O S S O COOH NH2 COOH NH2 S S S S S 70 80 90 mustard oil 10 (concentration of test compounds: 700µM) allicine 20 Anwar et al., J. Sulfur Chem. 2008 0 decadiene 30 AM DATTS DATS DADS DAS control S S S S S S Jacob, Natural Product Reports 2006 S S Allyl disulfide technical grade, 80% Diallylpolysulfanes and Caco-2 Cells: Inhibition of Proliferation S 100 S S 60 Cell Viability 50 [%] 40 S S S S Organic Sulfur Compounds: Plants and Fungi 19 17 18 Marut et al., Arthritis Research & Therapy, 2013 Dipropyltetrasulfane and Scleroderma Sulphur is an important detoxification for the liver and kidneys. It plays an energetic and anti-fatigue role as it preserves the energetic potential that is reactivated during a physical and intellectual effort. It also prevents muscle aches after strenuous physical efforts. Young people, Athletes and Intellectuals The demand of Sulphur is more important with its free antiradical actions, as the first signs of aging are numbness of the joints (often at night or when arising in the morning). The Elderly 20 Harlem oil provides a highly bioavailable sulfur ! Haarlem oil has demonstrated its effectiveness in preventing respiratory illness, passive and active smoking, unhealthy diets and rheumatoid arthritis. It is an energy supplement that promotes recovery after exercise, avoid aches and reduces mental fatigue. The Genuine Haarlem Oil is composed of three simple elements inside a 200mg capsule: •Sulphur 16% •Pine turpentine 80% •Linseed oil 4% Sulfur-based remedies: Haarlem oil 23 Thioaldehyde formation from α,β-unsaturated thiols Schneider et al., Med. Chem. Comm. 2011 Facets of the biological chemistry of polysulfanes 21 Czepukojc et al., Food and Chemical Toxicology 2013 Schematic overview of a prospective mode of action of the diallyl- and dipropyltetrasulfanes. 24 22 A.K. Baltes, Diploma thesis Microscopic analysis of influence of the tetrasulfanes on the tubulin network in U937 cells after 4 h incubation time. Polysulfanes trigger apoptosis in certain cells Benavides et. al. Natl . Acad. Sci. USA 2007 Das et. al. Am. J. Physiol. Heart. Circ. Physiol. 2007 Jacob et al., Planta Med. 2008 Simple H2S release from di-, tri- and tetrasulfanes The Special Role of Selenium and Tellurium Applications From Nutrition to Wider • „Green“ Pesticides and Fungicides 27 Agricultural Applications: Scleroderma Cancer Therapy Medical Applications: Possible applications of the chalcogen compounds 25 Slight inhibition 0.1 mM Less germination 0.3 mM Strong germination Concentration 0.2 mM Reactive Sulfur Species trigger apoptosis in certain cells Death 0.4 mM 28 Botrytis cinerea on tomato steam, Source: wildaboutbritain.co.uk Growth inhibition of MDR2 strain on Dithiolethione Botrytis cinerea on grape, Source: acenologia.com Czepukojc et al., Natural Product Communications 2013 Control MDR2 S S Dithiolethione S Botrytis assay 26 From Sulfur to Selenium and Tellurium Schneider et al., J. Biomed. Nanotechnol. 2011 Sulfur-based remedies: Nanoparticles 31 29 O NH N Se O N NH HN N SOD mimics: O N NH O Se IC50 5-10 µM Sensor/effector-agents (CLL leukemia): Water O O 4 O H N Note: There is a brand-new cluster of Se and Te chemists and biochemists emerging across Europe !! NC 3 O CO2H 2 1 IC50 5-10 µM Multicomponent Ugi-reaction: Sensor/effector agents Chalcogen-based remedies 32 30 33 Te Te 0 500 1000 1500 2000 J10 J12 Days J14 J17 Jamier et al., Chem. Eur. J. 2010 35 CT26 colon tumor Control P10 Oxaliplatine P10+oxaliplatine Catalysis as key to efficiency and selectivity Doering et al., J. Med. Chem. 2010 O O Coriat et al., Cell Death and Disease 2011 Apoptosis of CLL B-cells and PBMNC cells as measured by flow cytometry Cytotoxicity and Apoptosis of CLL B- and PBMNC cells Tumor Volume mm3 as Haemolysis of RBCs Seleninic acid reactive “warhead” Du et al., MedChemComm 2014 Antifungal activity Amphiphilic seleninic acids as ‘double impact’ agents Marut et al., J. Invest. Dermatol. 2012 Effects of Te compounds on skin fibrosis induced by HOCl in BALB/c mice 36 34 Mániková et al., Molecules 2014 Doering et al., Tetrahedron 2012 Chemogenetic analysis in yeast Deciphering the mode of action 39 37 18 h (a and d as negative controls) Schneider et al., MedChemComm 2012 structures (lower row) of PtK2 cells after incubation with Te compounds for Fluorescent microscopy of microtubuli (upper row) and microfilament Cellular staining techniques Schneider et al., MedChemComm 2012 Hierarchical cluster analysis of growth data measured by impedance: Tellurium compounds cluster with tubulysin B, griseofulvin and nocodazole 40 Impedance, cell growth curves and cluster analysis 38 43 context of inflammatory disease(s) Possible applications in the Schneider et al., MedChemComm 2012 interactions using KB-3-1 cell extracts and Pronase Drug Affinity Responsive Target Stability Analysis of compound-protein DARTS with Western Blot analysis 41 44 Doering et al.,Tetrahedron 2012 High efficiency of nanomolar concentrations of Te compounds against macrophages What’s next ?? 42 P. Arsenjans M. Chovanec N. Latruffe / M. Cherkaoui Malki G. Kirsch/ D. Bagrel (Fr) M. Diederich (Lux/Korea) F. Batteux (Fr) Ludger Wessjohann (Halle) Karl-Herbert Schäfer (Zweibrücken) M. Herling (Cologne) A. Slusarenko (Aachen) F. Sasse (Braunschweig) C.-M. Lehr (UdS) A. Kiemer (UdS) I. Bernhardt (UdS) M. Montenarh (UdS) Schlumberger Foundation German-French University BMBF DAAD Bruno Bock Thiochemical-S European Union (RedCat; Corena) Saarland (LFFP) Deutsche Forschungsgemeinschaft (DFG) Saarland University Money for nothin' and chicks for free Exotic Adventures 47 45 Marina Hakenesch Sandra Hübgen Javeria Iftikhar Nour Jamal Aldabbas Sharoon Griffin Dr. Uma M. Viswanathan Nassifatou Koko Tittikpina Lisa Faulstich Adel Al-Marby Jawad Nasim http://www.ijb.univlorraine.fr/spip.php?article176&lang=fr France Metz Technopôle 1 – 3 October 2014 Nutrition and Ageing Workshop NutriOx 2014 “NutriOx” 2014 in Metz Nashrawan Lababedi Dr. Ethiene Estevam 48 Aman Bhasin 46 Dr. Torsten Burkholz The Team: “Academiacs International”
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