Genomes and Genes
Summary: A non-heme iron-sulfur protein isolated from Clostridium pasteurianum and other bacteria. It is a component of NITROGENASE, which is active in nitrogen fixation, and consists of two subunits with molecular weights of 59.5 kDa and 50.7 kDa, respectively.
- Spatzal T, Aksoyoglu M, Zhang L, Andrade S, Schleicher E, Weber S, et al. Evidence for interstitial carbon in nitrogenase FeMo cofactor. Science. 2011;334:940 pubmed publisher..Atomic-resolution x-ray diffraction data and an electron spin echo envelope modulation (ESEEM) analysis now provide direct evidence that the ligand is a carbon species...
- Peters J, Stowell M, Soltis S, Finnegan M, Johnson M, Rees D. Redox-dependent structural changes in the nitrogenase P-cluster. Biochemistry. 1997;36:1181-7 pubmed..These observed redox-mediated structural changes of the P-cluster suggest a role for this cluster in coupling electron transfer and proton transfer in nitrogenase. ..
- Hu Y, Fay A, Lee C, Ribbe M. P-cluster maturation on nitrogenase MoFe protein. Proc Natl Acad Sci U S A. 2007;104:10424-9 pubmed..Furthermore, we provide spectroscopic evidence that the [4Fe-4S] cluster-like fragments can be converted to P-clusters, thereby firmly establishing the physiological relevance of the previously identified P-cluster precursor...
- Einsle O, Tezcan F, Andrade S, Schmid B, Yoshida M, Howard J, et al. Nitrogenase MoFe-protein at 1.16 A resolution: a central ligand in the FeMo-cofactor. Science. 2002;297:1696-700 pubmed..The presence of a nitrogen atom in the cofactor would have important implications for the mechanism of dinitrogen reduction by nitrogenase. ..
- Ribbe M, Burgess B. The chaperone GroEL is required for the final assembly of the molybdenum-iron protein of nitrogenase. Proc Natl Acad Sci U S A. 2001;98:5521-5 pubmed..The involvement of the molecular chaperone GroEL in the insertion of a metal cluster into an apoprotein may have broad implications for the maturation of other metalloenzymes. ..
- Rupnik K, Lee C, Hu Y, Ribbe M, Hales B. [4Fe4S]2+ clusters exhibit ground-state paramagnetism. J Am Chem Soc. 2011;133:6871-3 pubmed publisher..This unique characteristic suggests a specific coupling in these clusters necessary for nitrogen fixation and implies an evolutionary connection between the clusters in the two proteins. ..
- Goodwin P, Agar J, Roll J, Roberts G, Johnson M, Dean D. The Azotobacter vinelandii NifEN complex contains two identical [4Fe-4S] clusters. Biochemistry. 1998;37:10420-8 pubmed
- Hu Y, Corbett M, Fay A, Webber J, Hodgson K, Hedman B, et al. Nitrogenase Fe protein: A molybdate/homocitrate insertase. Proc Natl Acad Sci U S A. 2006;103:17125-30 pubmed
- Hu Y, Yoshizawa J, Fay A, Lee C, Wiig J, Ribbe M. Catalytic activities of NifEN: implications for nitrogenase evolution and mechanism. Proc Natl Acad Sci U S A. 2009;106:16962-6 pubmed publisher..More importantly, the discrepancy between the two systems may provide useful insights into nitrogenase mechanism and allow reconstruction of a fully functional nitrogenase from the "skeleton" enzyme, NifEN. ..
- Hu Y, Fay A, Ribbe M. Identification of a nitrogenase FeMo cofactor precursor on NifEN complex. Proc Natl Acad Sci U S A. 2005;102:3236-41 pubmed..The FeMoco-maturation assay described here will further address the remaining questions related to the assembly mechanism of the ever-intriguing FeMoco...
- Hoffman B, Dean D, Seefeldt L. Climbing nitrogenase: toward a mechanism of enzymatic nitrogen fixation. Acc Chem Res. 2009;42:609-19 pubmed publisher..Although the summit remains distant, we look forward to continued progress in the ascent. ..
- Broach R, Rupnik K, Hu Y, Fay A, Cotton M, Ribbe M, et al. Variable-temperature, variable-field magnetic circular dichroism spectroscopic study of the metal clusters in the DeltanifB and DeltanifH mofe proteins of nitrogenase from Azotobacter vinelandii. Biochemistry. 2006;45:15039-48 pubmed..These results suggest that the DeltanifH MoFe protein contains a pair of neighboring, unusual [4Fe-4S]-like clusters, which are paramagnetic in their oxidized state. ..
- Lancaster K, Roemelt M, Ettenhuber P, Hu Y, Ribbe M, Neese F, et al. X-ray emission spectroscopy evidences a central carbon in the nitrogenase iron-molybdenum cofactor. Science. 2011;334:974-7 pubmed publisher..Identification of the central atom will drive further studies on its role in catalysis. ..
- Ribbe M, Hu Y, Guo M, Schmid B, Burgess B. The FeMoco-deficient MoFe protein produced by a nifH deletion strain of Azotobacter vinelandii shows unusual P-cluster features. J Biol Chem. 2002;277:23469-76 pubmed..These unusual catalytic and spectroscopic properties might indicate the presence of a P-cluster precursor or a P-cluster trapped in an unusual conformation or oxidation state. ..
- Schmid B, Ribbe M, Einsle O, Yoshida M, Thomas L, Dean D, et al. Structure of a cofactor-deficient nitrogenase MoFe protein. Science. 2002;296:352-6 pubmed..A predominantly positively charged funnel is revealed; this funnel is of sufficient size to accommodate insertion of the negatively charged cofactor...
- Schmid B, Einsle O, Chiu H, Willing A, Yoshida M, Howard J, et al. Biochemical and structural characterization of the cross-linked complex of nitrogenase: comparison to the ADP-AlF4(-)-stabilized structure. Biochemistry. 2002;41:15557-65 pubmed
- Lee H, Igarashi R, Laryukhin M, Doan P, Dos Santos P, Dean D, et al. An organometallic intermediate during alkyne reduction by nitrogenase. J Am Chem Soc. 2004;126:9563-9 pubmed..Of the two most attractive structures, one singly reduced at C3 (4), the other being the doubly reduced allyl alcohol product (6), we tentatively favor 6 because of the "natural" assignment it affords for H(b). ..
- Mills P, Footitt E, Ceyhan S, Waters P, Jakobs C, Clayton P, et al. Urinary AASA excretion is elevated in patients with molybdenum cofactor deficiency and isolated sulphite oxidase deficiency. J Inherit Metab Dis. 2012;35:1031-6 pubmed publisher..This should be taken into account when interpreting the laboratory data. Sulphite was shown to inhibit ?-aminoadipic semialdehyde dehydrogenase in vitro. ..
- Afshar S, Johnson E, de Vries S, Schröder I. Properties of a thermostable nitrate reductase from the hyperthermophilic archaeon Pyrobaculum aerophilum. J Bacteriol. 2001;183:5491-5 pubmed..When incubated at 100 degrees C, the purified nitrate reductase had a half-life of 1.5 h. This study constitutes the first description of a nitrate reductase from a hyperthermophilic archaeon...
- Igarashi R, Dos Santos P, Niehaus W, Dance I, Dean D, Seefeldt L. Localization of a catalytic intermediate bound to the FeMo-cofactor of nitrogenase. J Biol Chem. 2004;279:34770-5 pubmed..Refinement of the binding mode and site was accomplished by the use of density functional and force field calculations pointing to an eta(2) coordination at Fe-6 of the FeMo-cofactor. ..
- Siemann S, Schneider K, Behrens K, Knöchel A, Klipp W, Muller A. FeMo cofactor biosynthesis in a nifE- mutant of Rhodobacter capsulatus. Eur J Biochem. 2001;268:1940-52 pubmed..The implications of our findings for the biosynthesis of the FeMoco will be discussed. ..
- Zhang Y, Mei T, Yang D, Zhang Y, Wang B, Qu J. Synthesis and reactivity of thiolate-bridged multi-iron complexes supported by cyclic (alkyl)(amino)carbene. Dalton Trans. 2017;46:15888-15896 pubmed publisher..Furthermore, complexes 3 and 4 exhibit good exchange reactivity toward the azide anion to give novel thiolate-bridged diiron complexes with two azido ligands in a trans arrangement...
- Chatelet C, Meyer J. Mapping the interaction of the [2Fe-2S] Clostridium pasteurianum ferredoxin with nitrogenase MoFe protein. Biochim Biophys Acta. 2001;1549:32-6 pubmed..The positions on the ferredoxin molecule of the residues interacting with the MoFe protein were determined using the recently elucidated crystal structure of the homologous [2Fe-2S] ferredoxin from Aquifex aeolicus. ..
- Chen X, Zhang W, Duncan J, Lee S. Iron-amide-sulfide and iron-imide-sulfide clusters: heteroligated core environments relevant to the nitrogenase FeMo cofactor. Inorg Chem. 2012;51:12891-904 pubmed publisher..Cluster 10 presents an [Fe(4)NS(3)] core framework virtually isometric with the isostructural [Fe(4)S(3)X] subunit of the FeMo cofactor, thus providing a synthetic structural representation for this portion of the cofactor core. ..
- Paustian T, Shah V, Roberts G. Apodinitrogenase: purification, association with a 20-kilodalton protein, and activation by the iron-molybdenum cofactor in the absence of dinitrogenase reductase. Biochemistry. 1990;29:3515-22 pubmed..This suggests that sulfhydryl(s) is (are) important for the FeMoco-activation reaction. ..
- Hu Y, Ribbe M. Biosynthesis of the iron-molybdenum cofactor of nitrogenase. J Biol Chem. 2013;288:13173-7 pubmed publisher..Here, we review the recent progress in the research area of M-cluster assembly, with an emphasis on our work that provides useful insights into the mechanistic details of this process. ..
- Rubio L, Ludden P. Biosynthesis of the iron-molybdenum cofactor of nitrogenase. Annu Rev Microbiol. 2008;62:93-111 pubmed publisher..This effort is starting to provide insights into the structures of the FeMo-co biosynthetic intermediates and into the biochemical details of FeMo-co synthesis. ..
- Kastner J, Blöchl P. Model for acetylene reduction by nitrogenase derived from density functional theory. Inorg Chem. 2005;44:4568-75 pubmed..Our proposed mechanism is consistent with experimentally observed stereoselectivity and the ability of C2H2 to suppress H2 production by nitrogenase. ..
- Cui Z, Dunford A, Durrant M, Henderson R, Smith B. Binding sites of nitrogenase: kinetic and theoretical studies of cyanide binding to extracted FeMo-cofactor derivatives. Inorg Chem. 2003;42:6252-64 pubmed
- Fisher K, Newton W, Lowe D. Electron paramagnetic resonance analysis of different Azotobacter vinelandii nitrogenase MoFe-protein conformations generated during enzyme turnover: evidence for S = 3/2 spin states from reduced MoFe-protein intermediates. Biochemistry. 2001;40:3333-9 pubmed..J., and Thorneley, R. N. F. (1984) Biochem. J. 224, 887-909] and propose that they arise from reduced states of the MoFe protein and reflect different conformations of the FeMo cofactor with different protonation states. ..
- Hu Y, Fay A, Schmid B, Makar B, Ribbe M. Molecular insights into nitrogenase FeMoco insertion: TRP-444 of MoFe protein alpha-subunit locks FeMoco in its binding site. J Biol Chem. 2006;281:30534-41 pubmed..Our results provide some of the initial molecular insights into the FeMoco insertion process and, moreover, have useful implications for the overall scheme of nitrogenase assembly. ..
- Hu Y, Corbett M, Fay A, Webber J, Hedman B, Hodgson K, et al. Nitrogenase reactivity with P-cluster variants. Proc Natl Acad Sci U S A. 2005;102:13825-30 pubmed
- Dance I. New insights into the reaction capabilities of His195 adjacent to the active site of nitrogenase. J Inorg Biochem. 2017;169:32-43 pubmed publisher..The accumulated results suggest that protonated His195 could be the agent for the first, most difficult, transfer of H to bound substrate N2. ..
- Reiss J, Hahnewald R. Molybdenum cofactor deficiency: Mutations in GPHN, MOCS1, and MOCS2. Hum Mutat. 2011;32:10-8 pubmed publisher..For type A deficiency an effective substitution therapy has been described recently. ..
- Dance I. The mechanism of nitrogenase. Computed details of the site and geometry of binding of alkyne and alkene substrates and intermediates. J Am Chem Soc. 2004;126:11852-63 pubmed..Alternative binding sites and geometries for ethyne and ethene, relevant to the wild-type protein, are described. This model defines the location and scene for detailed investigation of the mechanism of nitrogenase. ..
- Chow T, Tabita F. Reciprocal light-dark transcriptional control of nif and rbc expression and light-dependent posttranslational control of nitrogenase activity in Synechococcus sp. strain RF-1. J Bacteriol. 1994;176:6281-5 pubmed..By contrast, transcription of the rbcL gene, which encodes the catalytic subunit of the key enzyme of CO2 fixation (a light-dependent process), was enhanced in the light. ..
- Scott K, Rolfe B, Shine J. Biological nitrogen fixation: primary structure of the Rhizobium trifolii iron protein gene. DNA. 1983;2:149-55 pubmed..The nif structural genes are preceded by a DNA sequence which is repeated at least three times in the Rhizobium trifolii genome. ..
- Cotton M, Rupnik K, Broach R, Hu Y, Fay A, Ribbe M, et al. VTVH-MCD study of the Delta nifB Delta nifZ MoFe protein from Azotobacter vinelandii. J Am Chem Soc. 2009;131:4558-9 pubmed publisher..These results suggest that the MoFe protein is synthesized in a stepwise fashion where NifZ is specifically required for the formation of the second P-cluster. ..
- Rittle J, Peters J. Fe-N2/CO complexes that model a possible role for the interstitial C atom of FeMo-cofactor (FeMoco). Proc Natl Acad Sci U S A. 2013;110:15898-903 pubmed publisher..The data presented are contextualized in support of a hypothesis wherein modulation of a belt Fe-C interaction in the FeMo-cofactor facilitates substrate binding and reduction. ..
- Zhou Z, Yan W, Wan H, Tsai K. Synthesis and characterization of homochiral polymeric S-malato molybdate(VI): toward the potentially stereospecific formation and absolute configuration of iron-molybdenum cofactor in nitrogenase. J Inorg Biochem. 2002;90:137-43 pubmed..The absolute configuration of wild-type FeMo-cofactor is assigned as Delta(R). ..
- Vijayakumar K, Gunny R, Grunewald S, Carr L, Chong K, Devile C, et al. Clinical neuroimaging features and outcome in molybdenum cofactor deficiency. Pediatr Neurol. 2011;45:246-52 pubmed publisher..Imaging pattern on early brain MRI (<1 week) may prompt the diagnosis, potentially allowing early treatment and disease modifications. ..
- Pickett C, Vincent K, Ibrahim S, Gormal C, Smith B, Best S. Electron-transfer chemistry of the iron-molybdenum cofactor of nitrogenase: delocalized and localized reduced states of FeMoco which allow binding of carbon monoxide to iron and molybdenum. Chemistry. 2003;9:76-87 pubmed
- Hitzert M, Bos A, Bergman K, Veldman A, Schwarz G, Santamaria Araujo J, et al. Favorable outcome in a newborn with molybdenum cofactor type A deficiency treated with cPMP. Pediatrics. 2012;130:e1005-10 pubmed publisher..We strongly recommend starting cPMP treatment as soon as possible after birth in infants diagnosed with MoCD type A. ..
- Hinnemann B, Nørskov J. Modeling a central ligand in the nitrogenase FeMo cofactor. J Am Chem Soc. 2003;125:1466-7 pubmed..By comparison of bond geometries with the crystallographically determined values, we show that the central ligand is most likely nitrogen. ..
- Hales B. Magnetic circular dichroism spectroscopy. Methods Mol Biol. 2011;766:207-19 pubmed publisher..The basics of this technique are discussed along with examples of how MCD spectroscopy has been successfully used to elucidate the metal clusters of Nif proteins from nitrogen-fixing bacteria. ..
- Rupnik K, Lee C, Wiig J, Hu Y, Ribbe M, Hales B. Nonenzymatic synthesis of the P-cluster in the nitrogenase MoFe protein: evidence of the involvement of all-ferrous [Fe4S4](0) intermediates. Biochemistry. 2014;53:1108-16 pubmed publisher..These results demonstrate that the nitrogenase P-cluster can be generated in the absence of NifH and MgATP. ..
- Tolland J, Thorneley R. Stopped-flow Fourier transform infrared spectroscopy allows continuous monitoring of azide reduction, carbon monoxide inhibition, and ATP hydrolysis by nitrogenase. Biochemistry. 2005;44:9520-7 pubmed