SPP 1927:  Iron-Sulfur for Life

• Subject Area: Biology ; Chemistry ; Medicine
• Term: from 2016 to 2024
• Website: Homepage
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 273919336

Iron-sulfur (FeS) centers are essential protein cofactors in all forms of life. They are involved in many of the key biological processes including respiration, photosynthesis, metabolism of nitrogen, sulfur, carbon and hydrogen, biosynthesis of antibiotics, gene regulation, protein translation, replication and DNA repair, protection from oxidizing agents, and neurotransmission. In particular, FeS centers are not only involved as enzyme cofactors in catalysis and electron transfer, but they are also indispensable for the biosynthesis of complex metal-containing cofactors. A prominent example is represented by the family of radical/Sadenosylmethionine- dependent enzymes, which were discovered in 2001. Members of this family play essential roles in the biosynthesis of metal centers as complex as the iron-molybdenum cofactor (FeMoco) of nitrogenase, the molybdenum cofactor (Moco) of various molybdoenzymes, the active sites of [Fe-Fe]- and [Fe]-hydrogenases and the tetrapyrrole cofactors of hemes, corrins and chlorins. In spite of the recent fundamental breakthroughs in metalloenzyme research, it has become evident that studies on single enzymes has to be transformed into the broader context of a living cell where biosynthesis, function, and disassembly of these fascinating metal cofactors are coupled in a dynamic fashion. The various biosynthetic pathways were found to be tightly interconnected through a complex crosstalk mechanism that involves the dependence on the bio-availability of distinct metal ions, in particular molybdenum, iron, tungsten and zinc. The current lack of knowledge of such interaction networks is due to the sheer complexity of the metal cofactor biosynthesis with regard to both the (genetic) regulation and (chemical) metal center assembly. Recent pioneering technical developments allowed the detailed investigation of the assembly, biosynthesis and catalysis of FeS-dependent enzymes in a cellular context, opening up a new era in studying metalloenzymes. Such studies are not only important for understanding fundamental cellular processes but they are also a prerequisite for providing a comprehensive view of the complex biosynthesis and the catalytic mechanism of metalloenzymes that underlie metal-related human diseases. These key features of metalloenzymes can only be implemented in a cellular context. Further current topics are biotechnological applications such as alternative (bio)fuels and nitrogen fertilization, which are crucially linked to hydrogenase and nitrogenase, both of which are cornerstones of the proposed program. Understanding the crosstalk of metal ions on a cellular basis requires multidisciplinary cooperative approaches that span the entire range from molecular biology, inorganic chemistry, biochemistry, cell and mouse biology, and structural biology to theory and spectroscopy. This proposal aims to adopt a unique interdisciplinary approach to study novel enzyme mechanisms, innovative model complexes, and new biogenesis pathways in the physiological context of metalloenzymes in living organisms. Based on these new perceptions and the recent technical improvements, the researchers of the consortium will now be able to tackle the highly-demanding analysis of the assembly, biosynthesis, catalysis and relevance in disease of selected FeS-dependent metalloproteins. The unique grouping of scientists will greatly benefit from their profound combined knowledge in this field, and together they will aim to develop a comprehensive view of the interplay of metalloproteins in cellular systems. Within the framework of the proposed priority program (SPP) "Iron-Sulfur for Life – Cooperative function of Iron-Sulfur Centers in Assembly, Biosynthesis, Catalysis and Disease" an interdisciplinary consortium will be formed of 26 established and three female junior scientists that join forces to synergistically address the following fundamental areas: Assembly of FeS proteins as a starting point for versatile functionality Biosynthesis and crosstalk of complex metal cofactors by FeS proteins Catalysis and functions of complex FeS proteins for bio-applications Disease-relevant roles of FeS proteins in cellular metal homeostasis To address these topics adequately, novel interdisciplinary approaches will be combined along with methodological advancements. Proteins crucial for energy metabolism, H2-conversion, and nitrogen-cycling in Bacteria, Archaea and Eukarya including mammals, plants and fungi will be the focus of the anticipated studies. Given the prominent role of German researchers as an important driver of exciting new developments in the field of FeS-dependent proteins, we envisage that a coordinated SPP will further strengthen existing collaborations and will foster new joint efforts leading towards a novel understanding of cellular pathways in a general context. We see this as essential for maintaining and extending the leading role of German scientists in this rapidly developing field. We propose a core assembly of scientists for this SPP who are uniquely positioned and have been selected for the task at hand, because of their conceptual and technical leadership. Within the program, expertise, chosen experimental approaches and technologies will be exchanged and made available in a highly synergistic and interdisciplinary way, aiming at breaking the borders between disciplines, model organisms and studies in living organisms. We anticipate that within the six years of SPP funding the group will make substantial progress towards answering key questions on how complex FeS-dependent cofactors are assembled at a cellular level, and how the crosstalk between different assembly machineries in a metal-dependent manner is mediated. These studies have the future perspective for understanding the catalytic roles of essential enzymes in a cellular context, and therefore to help the profound understanding of FeS-related diseases in humans. Further, the program will lead to the development of future cellular systems for bioenergy production, fertilization and bio-applications.

• DFG Programme: Priority Programmes

Projects

• An in vitro system for the study of [NiFe]-hydrogenase maturation (Applicant Soboh, Basem )
• Assembly and Functional Role of the Iron-Sulfur Clusters in Respiratory Complex I (Applicant Friedrich, Thorsten )
• Assembly and Maturation of the Iron-Sulfur Clusters of Nitrogenases (Applicant Einsle, Oliver )
• Assembly of the membrane-bound FeS centre of the Na+-translocating NQR (Applicants Fritz, Günter ;  Fritz-Steuber, Julia )
• Characterization and reaction mechanism of the nitrogenase-like reductase CfbC/CfbD involved in cofactor F430 biosynthesis (Applicant Layer, Gunhild Monika )
• Comparative membrane metalloproteomics of respiring anaerobes (Applicant Adrian, Lorenz )
• CooC2/AcsF and Cfd1/Nbp35: maturation of complex Fe/S-clusters by MinD-type ATPases (Applicant Dobbek, Holger )
• Coordination Funds (Applicant Leimkühler, Silke )
• Correlation of three-dimensional and electronic structure of [FeFe] hydrogenases (Applicants Birrell, James ;  Span, Ingrid )
• Crosstalk of iron-sulfur cluster assembly, metal homeostasis and the biosynthesis of molybdoenzymes (Applicant Leimkühler, Silke )
• Deciphering the function of Fe/S cofactors with alternative cluster ligands: model studies using synthetic analogues (Applicant Meyer, Franc )
• Deciphering the stepwise cellular assembly and integration of the [FeFe]-hydrogenase H-cluster (Applicant Happe, Thomas )
• Diurnal Dynamics in the Biogenesis of Iron-Sulfur Cluster and other Metalloproteins of Plants in Dependence on Cluster Assembly (Applicant Krämer, Ute )
• Double-cubane iron-sulfur clusters: a new cofactor in biology (Applicant Dobbek, Holger )
• Elucidating the mechanisms of [4Fe-4S] cluster insertions into the cytosolic iron-sulfur protein assembly component Nar1 (Applicant Braymer, Ph.D., Joseph James )
• [Fe]-hydrogenase: biosynthesis of the FeGP cofactor and its catalytic function (Applicant Shima, Ph.D., Seigo )
• Fueling CO2-fixation by detoxifying CO, what are the secrets behind the electron-bifurcating hydrogenase/formate dehydrogenase from homoacetogens? (Applicant Wagner, Tristan )
• Functional analysis of the Fe-S cluster containing chloroplast J-domain proteins CDJ3-5 (Applicant Schroda, Michael )
• Interconnections of metallo enzyme synthesis machineries in Geobacter metallireducens: cofactor assembly and function of W-/FeS-containing BamB (Applicant Boll, Matthias )
• Iron-Sulfur Center Regulation and Crosstalk of two Radical SAM Modifiers by one Electron Transfer Protein in Yeast? (Applicant Schaffrath, Raffael )
• Iron-sulfur cluster electronic structural evolution and its contribution to diverse functionality (Applicants DeBeer, Ph.D., Serena ;  Neese, Frank )
• Iron-sulfur cofactors involved in metal center assembly and catalysis of hydrogenase (Applicants Lenz, Oliver ;  Zebger, Ingo )
• Mitochondrial and cytosolic maturation of human FeS-dependent molybdenum cofactor synthesis MOCS1A proteins and links between both cofactor pathways (Applicant Schwarz, Günter )
• Molecular determinants for cytosolic iron-sulfur cluster insertion (Applicant Pierik, Antonio J. )
• Molybdenum cofactor-biosynthesis and crosstalk to FeS metabolism in Neurospora crassa after ectopic expression of Moco biosynthesis step 1 proteins (Applicant Mendel, Ralf R. )
• Mössbauer spectroscopic methods to study iron-sulfur assembly, disassembly and catalysis (Applicant Schünemann, Volker )
• Nitrogenase-like Biosynthesis of (Bacterio)chlorophylls (Applicant Moser, Jürgen )
• Real-Time Spectroscopy for the Analysis of Gas-processing Metalloenzymes (Applicant Stripp, Sven )
• Role of the mitochondrial Bol1 and Bol3 proteins in iron-sulfur cluster delivery to diverse recipient proteins (Applicant Lill, Roland )
• Simulating the Coupling of Catalysis and Electron Transfer along Chains of FeS-Clusters in Various Enzymes (Applicant Ullmann, Matthias )
• Spatiotemporal Assembly and Function of the Modular Formate Hydrogenlyase Complex (Applicant Pinske, Constanze )
• Sulfido dithiolene complexes modeling molybdenum and tungsten dependent oxiodreductases - investigating synthesis, catalysis and biological activity to elucidate structural uncertainties, structure-function relationships and biosynthesis (Applicant Schulzke, Carola )
• The role of FeS-glutaredoxins and their cofactors in diseases (Applicants Berndt, Carsten ;  Lillig, Christopher Horst )
• Towards a mechanistic understanding of the role of the iron-sulphur cluster-containing HypD protein in diatomic ligand biosynthesis of [NiFe]-hydrogenases (Applicant Sawers, Gary )
• Tungsten-dependent aldehyde oxidoreductases: specific W-cofactor insertion and interactions with Fe-S clusters in catalysis and maturation (Applicant Heider, Johann )

• Spokesperson: Professorin Dr. Silke Leimkühler