Trypanosoma brucei has a complex life cycle during which its single mitochondrion is subjected to major metabolic and morphological changes. Rcsb pdb gene view nfs1 nfs1, cysteine desulfurase. Architectural features of human mitochondrial cysteine. Although several functional studies support the physiological dependence of nfs1 for being active only with isd11, less attention has been given to investigate the molecular properties of each protein alone and in a complex and also in elucidating. Nfs1 is a desulfurase that acts in the abstraction of sulfur from cysteine for the formation of fes clusters in the mitochondria lill and muhlenhoff, 2005. Mitochondrial and nucleolar localization of cysteine. Mechanism of activation of the human cysteine desulfurase. Download dna or protein sequence, view genomic context and coordinates.
The nfs1isd11acp complex is the central subcomplex involved in providing sulfur for cluster synthesis. Learn vocabulary, terms, and more with flashcards, games, and other study tools. This message will disappear when all data is loaded. Both nfs1 and iscs generate a persulfide intermediate on a conserved cysteine residue of a mobile stransfer loop. Mitochondrial cysteine desulfurase and isd11 coexpressed.
Nfs1 nfs1, cysteine desulfurase gene view the gene browser allows to navigate the human genome and investigate the relationship between pdb entries and genes. Sep 24, 2019 in 2017, the first crystal structure of a mitochondrial cysteine desulfurase sda ec complex surprisingly revealed an incomplete substratebinding site, a solventexposed plp cofactor, a disordered mobile stransfer loop, and an overall open. Human isd11 is essential for both ironsulfur cluster. Little is known regarding the dynamics of the interaction of proteins with the fes cluster scaffold isu1. Cysteine desulfurase an overview sciencedirect topics. Download dna or protein sequence, view genomic context. The data was collected at ssrl bl 71 and published in pnas june 2017. Gene references into functions our findings highlight that the isd11 r68ar68l mutation display reduced affinity to form a stable subcomplex with nfs1, and thereby fails to.
Assembly of fes clusters requires cysteine desulfurases, such as nfs1, as well as scaffold proteins, iron donors, and chaperones li et al. Pdf nfs1 cysteine desulfurase protein complexes and. The lcysteine desulfurase nfs1 is localized in the cytosol where it provides the sulfur for molybdenum cofactor biosynthesis in humans. In humans, the l cysteine desulfurase nfs1 plays a crucial role in the mitochondrial ironsulfur cluster biosynthesis and in the thiomodification of mitochondrial and cytosolic trnas. This gene uses alternate inframe translation initiation sites to generate. The nfs1 interacting protein isd11 has an essential role. Mitochondria exported these species via the atm1 transporter in the inner membrane. Thus, the two substrates of this enzyme are lcysteine and enzymecysteine, whereas its two products are lalanine and enzymessulfanylcysteine. Mitochondria were found to utilize the cysteine desulfurase nfs1 to produce sulfurcontaining species with masses ranging from 700 to 1,100 da. Increased evidence for the functions of cysteine desulfurases has revealed their important. Frataxin directly stimulates mitochondrial cysteine desulfurase by exposing substratebinding sites, and a mutant fes cluster scaffold. The ferrous sulfate will react with the hydrogen sulfide gas and produce ferrous sulfide, which has a dark black appearance.
Interpro provides functional analysis of proteins by classifying them into families and predicting domains and important sites. In this study, we report that expression of two essential components of the fes machinery, the cysteine desulfurase nfs1 and its scaffold protein partner iscu, is downregulated at both mrna and protein levels when murine macrophages are physiologically stimulated with ifn. Characterization of yeast cysteine desulfurase complex and. In humans, the lcysteine desulfurase nfs1 is the orthologue of azotobacter vinelandii nifs and escherichia coli iscs. We combine protein signatures from a number of member databases into a single searchable resource, capitalising on their individual strengths to produce a powerful integrated database and diagnostic tool. Gene ontology go annotations related to this gene include protein. Involved in the 2thiomodification of both 5carboxymethylaminomethyl2thiouridine in mitochondrial trnas and 5. Additionally, we analyzed the localization of isd11, the partner protein of nfs1 in mitochondria. Cysteine desulfurase nfs1 and pim1 protease control levels of isu, the fes cluster biogenesis scaffold jiyoon songa, jaroslaw marszaleka,b, and elizabeth anne craiga,1 adepartment of biochemistry, university of wisconsin, madison, wi 53706. Fxn accelerates the persulfide formation kinetics on both nfs1 and iscu2. In humans, the lcysteine desulfurase nfs1 plays a crucial role in the mitochondrial ironsulfur cluster biosynthesis and in the thiomodification of mitochondrial and cytosolic trnas. Eukaryotic nfs1 tends to aggregate unless its small partner protein isd11 is bound. The l cysteine desulfurase nfs1 is localized in the cytosol where it provides the sulfur for molybdenum cofactor biosynthesis in humans. The lcysteine desulfurase nfs1 is localized in the cytosol.
Lcysteine desulfurases are pyridoxal phosphate plpdependent enzymes that use lcysteine as substrate to produce lalanine and a proteinbound persulfide. Cysteine desulfurase, mitochondrial is an enzyme that in humans is encoded by the nfs1 gene ironsulfur clusters are required for the function of many cellular enzymes. While the procyclic stage ps of the insect vector contains a large and reticulated mitochondrion, its counterpart in the bloodstream stage bs parasitizing mammals is highly reduced and seems to be devoid of. B3kma5, b4dxk9, e1p5r8, f5gyk5, q6p0l8, q9ntz5, q9y481 this subsection of the entry information section shows the date of integration of the entry into uniprotkb, the date of the last sequence update and the date of the last annotation modification last. Frataxin directly stimulates mitochondrial cysteine. It supplies the inorganic sulfur for iron sulfur fes clusters. Neither the content nor the benchsci technology and processes for selection have been evaluated by us. Download citation cysteine desulfurase nfs1 and pim1 protease control levels of isu, the fes cluster biogenesis scaffold fes clusters are critical prosthetic groups for proteins involved in. My data nfs1 cysteine desulfurase, mitochondrial in homo sapiens. The structure revealed that nfs1 forms an unexpected cysteine desulfurase dimer architecture that is stabilized by the 3helix adaptor protein isd11. Increased evidence for the functions of cysteine desulfurases has revealed their important roles in the biosyntheses of fes. The lcysteine desulfurase nfs1 is localized in the cytosol where. The key steps involve mobilization of sulfur from cysteine by a cysteine desulfurase nfs1isd11 for the yeast proteins, delivery of iron, formation of an fes cluster intermediate on the scaffold proteins isu1 and isu2, and subsequent transfer of the cluster intermediates to apoproteins via a chaperone system lill et al.
In yeast, the cysteine desulfurase nfs1 is supported by the accessory protein isd11. Physiologically relevant reconstitution of ironsulfur. Users can perform simple and advanced searches based on. Nfs1 cysteine desulfurase protein complexes and phosphorylation sites as assessed by mass spectrometry. Sasddb3 human mitochondrial cysteine desulfurase nfs1. Cysteine desulfurase activities and in vitro fes cluster assembly rates of different reaction mixtures. Using antibodies to isd11 and iscs, we demonstrated complex formation between endogenous isd11 and iscs in vivo with coimmunoprecipitation assays fig. May 17, 2019 the nfs1 cysteine desulfurase facilitates a pyridoxal 5. The nfs1 interacting protein isd11 has an essential role in.
Cysteine desulfurase is regulated by phosphorylation of. Nfs1 nfs1 cysteine desulfurase is a protein coding gene. The rcsb pdb also provides a variety of tools and resources. Plays a role in both trnaprocessing and mitochondrial metabolism. Three conserved isu1 residues are critical for interaction with cysteine desulfurase nfs1 and jprotein cochaperone jac1, required for cluster assembly and transfer, respectively. C had similar cysteine desulfurase activity to wt nfs1 42. We describe results from chemical crosslinking coupled with tandem mass spectrometry and smallangle xray scattering studies that are consistent with a closed nfs1 dimer rather than an open one for both the cysteine. Cysteine desulfurase plays a central role in mitochondrial ironsulfur cluster. Diseases associated with nfs1 include combined oxidative phosphorylation deficiency 19 and xanthinuria. The lcysteine desulfurase nfs1 is localized in the.
The enzyme is involved in the biosynthesis of ironsulfur clusters, thionucleosides in trna, thiamine, biotin, lipoate and pyranopterin molybdopterin. It supplies the inorganic sulfur for ironsulfur fes clusters. Mitochondrial cysteine desulfurase is an essential component of the machinery for ironsulfur cluster biosynthesis. The j biological c 2004 by the american society for. Catalyzes the removal of elemental sulfur from cysteine to produce alanine. Binding of the chaperone jac1 protein and cysteine. The role of the accessory protein isd11 within the catalytic role of the cysteine desulfurase nfs1 in delivering sulfane sulfur for cluster assembly has been studied.
The nfs1 cysteine desulfurase facilitates a pyridoxal 5. Mitochondria export sulfur species required for cytosolic. The ferredoxin yah1 and ferredoxin reductase arh1 supply the required electrons 8. Thus, eukaryotic nfs1 is a functional orthologue of the bacterial cysteine desulfurase iscs. We tested the desulfurase activity in mitochondrial extracts from cells depleted of isd11 and from wt cells.
Cysteine desulfurase nfs1 is required generation of a. The effect of yfh1 on sulfur mobilization by the nfs1isd11 complex has also been evaluated in detail. We have previously demonstrated that purified nfs1 is able to transfer sulfur to the cterminal domain of mocs3, a cytosolic protein involved in molybdenum cofactor biosynthesis and trna thiolation. Discrete chemical steps in the first half of the cysteine desulfurase reaction involving the binding of cysteine and. Ironsulfur clusters are required for the function of many cellular. Ironsulfur clusters are required for the function of many cellular enzymes. Characterization of yeast cysteine desulfurase complex. Cysteine desulfurase is regulated by phosphorylation of nfs1. Fes clusters are biosynthesized on the scaffold protein iscu, with cysteine desulfurase nfs1 providing sulfur as persulfide and ferredoxin fdx2 supplying electrons, in a process stimulated by fxn. Stuart3, jan tachezy4, marinus pilon5 and julius lukes. The pdb archive contains information about experimentallydetermined structures of proteins, nucleic acids, and complex assemblies. However, less isu was coimmunoprecipitated from mitochondrial lysates with nfs1. Involved in the 2thiomodification of both 5carboxymethylaminomethyl2thiouridine in mitochondrial trnas and 5methoxycarbonylmethyl2thiouridine mcm5s2u in cytoplasmic trnas.
Characterization of the yeast cysteine desulfurase. Nfs1 2 with either refdx1 or refdx2 as the reducing agent. In yeast, fes cluster assembly on isu1 critically depends on the function of the cysteine desulfurase complex nfs1isd11 47, 54, 55. Nfs1 cysteine desulfurase, mitochondrial precursor. Sgd the effect of the adaptor protein isd11 on the quaternary structure of the eukaryotic cysteine desulphurase nfs1. Comments the sulfur from free lcysteine is first transferred to a cysteine residue in the active site, and then passed on to various other acceptors. Neither the content nor the benchsci technology and. We have previously demonstrated that purified nfs1 is able to transfer sulfur to the cterminal domain of mocs3, a cytosolic protein involved in molybdenum. Mitochondrial cysteine desulfurase and isd11 coexpressed in. Human mitochondrial ferredoxin 1 fdx1 and ferredoxin 2. Yfh1, the yeast frataxin homolog, has been proposed to serve as an iron donor andor a regulator of nfs1 function 911.
Cysteine desulfurase, mitochondrial is an enzyme that in humans is encoded by the nfs1. Once exported to the cytosol, these sulfur species promoted cytosolic trna thiolation with no further requirement of. As a member of the wwpdb, the rcsb pdb curates and annotates pdb data according to agreed upon standards. Cysteine desulfurase nfs1 and pim1 protease control levels of. Cysteine desulfurase nfs1 and pim1 protease control levels. Desulfurase definition of desulfurase by medical dictionary. Pdf the lcysteine desulfurase nfs1 is localized in the.
Thus, the two substrates of this enzyme are lcysteine and enzymecysteine, whereas its two products are lalanine and enzymessulfanylcysteine this enzyme belongs to the family of transferases. The human mitochondrial cysteine desulfurase complex consists of two copies each of nfs1, isd11, and acyl carrier protein. Saxs data from solutions of human mitochondrial cysteine desulfurase nfs1, isd11 and acp heterodimer complex in 20 mm hepes, 150 mm nacl, 5 mm tcep, ph 7. Most relevant to this report, nfs1, a 51kda cysteine desulfurase, donates the sulfur needed for fes cluster synthesis from cysteine, transferring it to isu. The proteins encoded by this gene supply inorganic sulfur to these clusters by removing the sulfur from cysteine, creating alanine in the process. This indicates that the enzyme cysteine desulfhydrase is present because it has produced h.
Users can perform simple and advanced searches based on annotations relating to sequence. As isd11 and the cysteine desulfurase, nfs1, were recently shown to form a complex in s. Regulation of the cysteine desulfurase nfs1 and the. An enzyme that catalyzes the removal of sulfur, usually as hydrogen sulfide, from organic compounds. The sulfur from free l cysteine is first transferred to a cysteine residue in the active site, and then passed on to various other acceptors. Clicking the images or links will redirect you to a website hosted by benchsci that provides thirdparty scientific content. Structure of the human cysteine desulfurase complex. Functions and cellular localization of cysteine desulfurase and selenocysteine lyase in trypanosoma brucei pavel poliak1, douglas van hoewyk2, miroslav oborn. Among its related pathways are metabolism of watersoluble vitamins and cofactors and mitochondrial ironsulfur cluster biogenesis. May be involved in the biosynthesis of molybdenum cofactor 457 aa. Second, we showed that only the mitochondrial version but not the extramitochondrial version of nfs1p is functional in generating cytosolic and nuclear fes proteins.
It has long been thought that nfs1 alone suffices to liberate sulfur from cysteine because the purified recombinant protein contains the enzymatic activity as a cysteine desulfurase and releases sulfide in. Nfs1 is mainly localized in mitochondria where it acts as a component of the. Antibodies for proteins involved in cysteine desulfurase activity pathways. Nfs1 cysteine desulfurase, mitochondrial precursor homo. It has been known that human cysteine desulfurase that is catalytically active in vitro can be prepared by overexpressing in escherichia coli cells two protein components of this system, the cysteine desulfurase protein nfs1 and the auxiliary protein isd11. While the procyclic stage ps of the insect vector contains a large and reticulated mitochondrion, its counterpart in the bloodstream stage bs parasitizing mammals is highly reduced and seems to be devoid of most functions. The sdaecu and sdaecuf complexes were reacted with a. In 2017, the first crystal structure of a mitochondrial cysteine desulfurase sda ec complex surprisingly revealed an incomplete substratebinding site, a solventexposed plp cofactor, a disordered mobile stransfer loop, and an overall open.
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