EzCatDB: M00199
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DB codeM00199
RLCP classification10.22025.100.10765 : Electron transfer
1.15.8230.352 : Hydrolysis
10.25130.110.10590 : Electron transfer
CATH domainDomain 13.40.50.1980 : Rossmann foldCatalytic domain
Domain 23.40.50.1980 : Rossmann foldCatalytic domain
Domain 33.40.50.1980 : Rossmann foldCatalytic domain
Domain 43.40.50.1980 : Rossmann foldCatalytic domain
Domain 53.40.50.1980 : Rossmann fold
Domain 63.40.50.1980 : Rossmann fold
Domain 71.20.89.10 : Nitrogenase Molybdenum-iron Protein, subunit B; domain 4
Domain 83.40.50.300 : Rossmann foldCatalytic domain
E.C.1.18.6.1

CATH domainRelated DB codes (homologues)
1.20.89.10 : Nitrogenase Molybdenum-iron Protein, subunit B; domain 4M00114
3.40.50.1980 : Rossmann foldM00114
3.40.50.300 : Rossmann foldS00527,S00547,S00548,S00550,S00554,S00555,S00671,S00672,S00676,S00680,S00682,S00913,S00914,S00301,S00302,S00303,S00304,S00307,S00308,S00305,S00306,S00309,S00310,S00311,M00114,D00129,D00130,D00540,M00186

Enzyme Name
UniProtKBKEGG

P00467P11347P00456
Protein nameNitrogenase molybdenum-iron protein alpha chainNitrogenase molybdenum-iron protein beta chainNitrogenase iron protein 1nitrogenase
SynonymsEC 1.18.6.1
Nitrogenase component I
Dinitrogenase
EC 1.18.6.1
Nitrogenase component I
Dinitrogenase
EC 1.18.6.1
Nitrogenase component II
Nitrogenase Fe protein 1
Nitrogenase reductase
PfamPF00148 (Oxidored_nitro)
[Graphical view]
PF00148 (Oxidored_nitro)
[Graphical view]
PF00142 (Fer4_NifH)
[Graphical view]

KEGG pathways
MAP codePathways
MAP00625Tetrachloroethene degradation
MAP00910Nitrogen metabolism

UniProtKB:Accession NumberP00467P11347P00456
Entry nameNIFD_CLOPANIFK_CLOPANIFH1_CLOPA
Activity8 reduced ferredoxin + 8 H(+) + N(2) + 16 ATP + 16 H(2)O = 8 oxidized ferredoxin + H(2) + 2 NH(3) + 16 ADP + 16 phosphate.8 reduced ferredoxin + 8 H(+) + N(2) + 16 ATP + 16 H(2)O = 8 oxidized ferredoxin + H(2) + 2 NH(3) + 16 ADP + 16 phosphate.8 reduced ferredoxin + 8 H(+) + N(2) + 16 ATP + 16 H(2)O = 8 oxidized ferredoxin + H(2) + 2 NH(3) + 16 ADP + 16 phosphate.
SubunitTetramer of two alpha and two beta chains. Forms complex with the iron protein (nitrogenase component 2).Tetramer of two alpha and two beta chains. Forms complex with the iron protein (nitrogenase component 2).Homodimer.
Subcellular location


CofactorBinds 1 8Fe-7S cluster per heterodimer (By similarity).,Binds 1 7Fe-Mo-9S-X-homocitryl cluster per subunit. The identity of the X atom is not known, possibly carbon or oxygen (By similarity).Binds 1 8Fe-7S cluster per heterodimer.Binds 1 4Fe-4S cluster per dimer.

Compound table: links to PDB-related databases & PoSSuM

CofactorsSubstratesProductsintermediates
KEGG-idL00026L00025C01251L00024C00305C00138C00080C00697C00002C00001C00139C00014C00008C00009

Compound[8Fe-7S](P cluster)[7Fe-9S-Mo](FeMoco)Homocitrate[4Fe-4S]MagnesiumReduced ferredoxinH+N2ATPH2OOxidized ferredoxinNH3ADPOrthophosphateTransition-state for ATP hydrolysisIntermediate for Nitrogenation
Typeheavy metal,sulfide groupheavy metal,sulfide groupcarbohydrate,carboxyl groupheavy metal,sulfide groupdivalent metal (Ca2+, Mg2+)heavy metal,peptide/protein,sulfide groupothersothersamine group,nucleotideH2Oheavy metal,peptide/protein,sulfide groupamine group,organic ionamine group,nucleotidephosphate group/phosphate ion

ChEBI
48796
52222
33725
18420

15378
17997
15422
15377

16134
16761
26078


PubChem

439459

888

1038
947
5957
962
22247451

222
6022
22486802
1004


                        
1mioA01UnboundBound:CFMBound:HCAUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioC01UnboundBound:CFMBound:HCAUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioA02UnboundUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioC02UnboundUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioA03UnboundUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioC03UnboundUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioB01Analogue:CLPUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioD01Analogue:CLPUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioB02UnboundUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioD02UnboundUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioB04UnboundUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioD04UnboundUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioB03UnboundUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1mioD03UnboundUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1cp2AUnboundUnboundUnboundBound:SF4UnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 
1cp2BUnboundUnboundUnboundUnboundUnboundUnbound UnboundUnbound UnboundUnboundUnboundUnboundUnbound 

Active-site residues
pdbCatalytic residuesCofactor-binding residuesMain-chain involved in catalysis
           
1mioA01CYS 53
CYS 53(P cluster);ILE 465;HIS 482(Homocitrate);HIS 482(FeMoco)
 
1mioC01CYS 53
CYS 53(P cluster);ILE 465;HIS 482(Homocitrate);HIS 482(FeMoco)
 
1mioA02GLN 182
GLN 182(Homocitrate);CYS 79;CYS 145(P cluster)
 
1mioC02GLN 182
GLN 182(Homocitrate);CYS 79;CYS 145(P cluster)
 
1mioA03 
CYS 262(FeMoco)
 
1mioC03 
CYS 262(FeMoco)
 
1mioB01CYS 106;THR 110
CYS 23;CYS 48;CYS 106;SER 141(P cluster)
 
1mioD01CYS 106;THR 110
CYS 23;CYS 48;CYS 106;SER 141(P cluster)
 
1mioB02 
 
 
1mioD02 
 
 
1mioB04 
 
 
1mioD04 
 
 
1mioB03 
 
 
1mioD03 
 
 
1cp2ALYS  9;LYS 14;ASP 38;LYS 40;ASP 126
CYS 94;CYS 129(4Fe-4S binding);SER 15(Magnesium binding)
GLY 11
1cp2BLYS  9;LYS 14;ASP 38;LYS 40;ASP 126
CYS 94;CYS 129(4Fe-4S binding);SER 15(Magnesium binding)
GLY 11

References for Catalytic Mechanism
ReferencesSectionsNo. of steps in catalysis
[5]Fig.2
[12]Scheme 1, Scheme 2, Scheme 3
[14]p.1680-1681
[16]p.1657-1658
[17]p.994
[18]Fig.9, p.347-349
[20]p.7112-7113
[24]Fig.2, Fig.3, p.794
[33]p.27012-27013
[35]p.10722
[36]p.13115-13117
[40]p.16774-16775
[41]p.9431-9432
[42]p.15660-15661
[46]Scheme 1, p.12981-12982
[47]Fig.2, Fig.4
[50]p.374-375
[52]p.11381-11383
[53]p.404-406
[64]p.881-886
[68]p.14750-14751
[71]p.563-564
[73]Scheme 1, Scheme 2, Fig.3, Fig.4, p.571-575
[74]p.13824
[75]p.648-650
[78]

[81]Fig.6
[86]Fig.7, Fig.8
[87]Fig.5, p.1698-1699
[89]Scheme 1, Scheme 2, Scheme 3, Scheme 4
[91]p.1032-1033
[93]p.380-381
[94]Fig.3
[95]Fig.8, p.1048-1051

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Related PDB1nip
Related UniProtKBP00459
[17]
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Year1992
Volume258
Pages992-5
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Related PDB1mio
Related UniProtKBP00467,P11347
[21]
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Year1993
Volume32
Pages6058-64
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Year1995
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Year1996
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Year1996
Volume35
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Year1996
Volume35
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Year1996
Volume35
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Related PDB1min,2min,3min
Related UniProtKBP07328,P07329
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Year1997
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Year1997
Volume387
Pages370-6
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Related PDB1n2c
Related UniProtKBP07328,P07329
[51]
PubMed ID9730834
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Year1998
Volume37
Pages12611-23
AuthorsChristiansen J, Goodwin PJ, Lanzilotta WN, Seefeldt LC, Dean DR
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JournalBiochemistry
Year1998
Volume37
Pages11376-84
AuthorsLanzilotta WN, Christiansen J, Dean DR, Seefeldt LC
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Year1998
Volume37
Pages399-407
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TitleElectron transfer in nitrogenase analyzed by Marcus theory: evidence for gating by MgATP.
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Year1998
Volume432
Pages55-8
AuthorsSpee JH, Arendsen AF, Wassink H, Marritt SJ, Hagen WR, Haaker H
TitleRedox properties and electron paramagnetic resonance spectroscopy of the transition state complex of Azotobacter vinelandii nitrogenase.
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PubMed ID9756863
JournalJ Biol Chem
Year1998
Volume273
Pages26330-7
AuthorsAngove HC, Yoo SJ, Munck E, Burgess BK
TitleAn all-ferrous state of the Fe protein of nitrogenase. Interaction with nucleotides and electron transfer to the MoFe protein.
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PubMed ID9792679
JournalJ Biol Chem
Year1998
Volume273
Pages29678-85
AuthorsBursey EH, Burgess BK
TitleThe role of methionine 156 in cross-subunit nucleotide interactions in the iron protein of nitrogenase.
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PubMed ID9642255
JournalJ Biol Chem
Year1998
Volume273
Pages16927-34
AuthorsBursey EH, Burgess BK
TitleCharacterization of a variant iron protein of nitrogenase that is impaired in its ability to adopt the MgATP-induced conformational change.
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PubMed ID9677296
JournalJ Mol Biol
Year1998
Volume280
Pages669-85
AuthorsSchlessman JL, Woo D, Joshua-Tor L, Howard JB, Rees DC
TitleConformational variability in structures of the nitrogenase iron proteins from Azotobacter vinelandii and Clostridium pasteurianum.
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PubMed ID9860849
JournalBiochemistry
Year1998
Volume37
Pages17345-54
AuthorsDuyvis MG, Wassink H, Haaker H
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PubMed ID10215587
JournalBiochem J
Year1999
Volume339
Pages511-5
AuthorsYousafzai FK, Eady RR
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[61]
PubMed ID10231529
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Year1999
Volume38
Pages5779-85
AuthorsChan JM, Christiansen J, Dean DR, Seefeldt LC
TitleSpectroscopic evidence for changes in the redox state of the nitrogenase P-cluster during turnover.
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PubMed ID10364195
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Year1999
Volume274
Pages17593-8
AuthorsChan JM, Ryle MJ, Seefeldt LC
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[63]
PubMed ID10391920
JournalJ Biol Chem
Year1999
Volume274
Pages19778-84
AuthorsRangaraj P, Ryle MJ, Lanzilotta WN, Ludden PW, Shah VK
TitleIn vitro biosynthesis of iron-molybdenum cofactor and maturation of the nif-encoded apodinitrogenase. Effect of substitution for NifH with site-specifically altered forms of NifH.
[64]
CommentsX-ray crystallography
PubMed ID10525412
JournalJ Mol Biol
Year1999
Volume292
Pages871-91
AuthorsMayer SM, Lawson DM, Gormal CA, Roe SM, Smith BE
TitleNew insights into structure-function relationships in nitrogenase: A 1.6 A resolution X-ray crystallographic study of Klebsiella pneumoniae MoFe-protein.
Related PDB1qgu,1qh1,1qh8
[65]
PubMed ID10852721
JournalBiochemistry
Year2000
Volume39
Pages7221-8
AuthorsChan JM, Wu W, Dean DR, Seefeldt LC
TitleConstruction and characterization of a heterodimeric iron protein: defining roles for adenosine triphosphate in nitrogenase catalysis.
[66]
PubMed ID10985789
JournalBiochemistry
Year2000
Volume39
Pages11434-40
AuthorsClarke TA, Maritano S, Eady RR
TitleFormation of a tight 1:1 complex of Clostridium pasteurianum Fe protein-Azotobacter vinelandii MoFe protein: evidence for long-range interactions between the Fe protein binding sites during catalytic hydrogen evolution.
[67]
PubMed ID11087939
JournalBiochim Biophys Acta
Year2000
Volume1543
Pages36-46
AuthorsJohnson JL, Nyborg AC, Wilson PE, Tolley AM, Nordmeyer FR, Watt GD
TitleMechanistic interpretation of the dilution effect for Azotobacter vinelandii and Clostridium pasteurianum nitrogenase catalysis.
[68]
CommentsX-RAY CRYSTALLOGRAPHY (2.15 ANGSTROMS).
Medline ID20552920
PubMed ID11101289
JournalBiochemistry
Year2000
Volume39
Pages14745-52
AuthorsJang SB, Seefeldt LC, Peters JW
TitleInsights into nucleotide signal transduction in nitrogenase: structure of an iron protein with MgADP bound.
Related PDB1fp6
Related UniProtKBP00459
[69]
PubMed ID10651628
JournalBiochemistry
Year2000
Volume39
Pages641-8
AuthorsJang SB, Seefeldt LC, Peters JW
TitleModulating the midpoint potential of the [4Fe-4S] cluster of the nitrogenase Fe protein.
[70]
PubMed ID11087938
JournalBiochim Biophys Acta
Year2000
Volume1543
Pages24-35
AuthorsJohnson JL, Nyborg AC, Wilson PE, Tolley AM, Nordmeyer FR, Watt GD
TitleAnalysis of steady state Fe and MoFe protein interactions during nitrogenase catalysis.
[71]
PubMed ID11006545
JournalCurr Opin Chem Biol
Year2000
Volume4
Pages559-66
AuthorsRees DC, Howard JB
TitleNitrogenase: standing at the crossroads.
[72]
PubMed ID10837496
JournalJ Biol Chem
Year2000
Volume275
Pages17631-8
AuthorsRibbe MW, Bursey EH, Burgess BK
TitleIdentification of an Fe protein residue (Glu146) of Azotobacter vinelandii nitrogenase that is specifically involved in FeMo cofactor insertion.
[73]
PubMed ID11311117
JournalBiochem J
Year2001
Volume355
Pages569-76
AuthorsDurrant MC
TitleControlled protonation of iron-molybdenum cofactor by nitrogenase: a structural and theoretical analysis.
[74]
PubMed ID11705370
JournalBiochemistry
Year2001
Volume40
Pages13816-25
AuthorsBenton PM, Mayer SM, Shao J, Hoffman BM, Dean DR, Seefeldt LC
TitleInteraction of acetylene and cyanide with the resting state of nitrogenase alpha-96-substituted MoFe proteins.
[75]
CommentsX-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS)
Medline ID21093028
PubMed ID11170380
JournalBiochemistry
Year2001
Volume40
Pages641-50
AuthorsChiu H, Peters JW, Lanzilotta WN, Ryle MJ, Seefeldt LC, Howard JB, Rees DC
TitleMgATP-Bound and nucleotide-free structures of a nitrogenase protein complex between the Leu 127 Delta-Fe-protein and the MoFe-protein.
Related PDB1g20,1g21
Related UniProtKBP07328,P07329
[76]
PubMed ID11258953
JournalBiochemistry
Year2001
Volume40
Pages3333-9
AuthorsFisher K, Newton WE, Lowe DJ
TitleElectron 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.
[77]
CommentsX-ray crystallography
PubMed ID11327812
JournalBiochemistry
Year2001
Volume40
Pages1540-9
AuthorsSorlie M, Christiansen J, Lemon BJ, Peters JW, Dean DR, Hales BJ
TitleMechanistic features and structure of the nitrogenase alpha-Gln195 MoFe protein.
Related PDB1fp4
[78]
PubMed ID11734043
JournalJ Am Chem Soc
Year2001
Volume123
Pages12392-410
AuthorsLovell T, Li J, Liu T, Case DA, Noodleman L
TitleFeMo cofactor of nitrogenase: a density functional study of states M(N), M(OX), M(R), and M(I).
[79]
CommentsX-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS).
Medline ID21093029
PubMed ID11170381
JournalBiochemistry
Year2001
Volume40
Pages651-6
AuthorsStrop P, Takahara PM, Chiu H, Angove HC, Burgess BK, Rees DC
TitleCrystal structure of the all-ferrous [4Fe-4S]0 form of the nitrogenase iron protein from Azotobacter vinelandii.
Related PDB1g1m,1g5p
Related UniProtKBP00459
[80]
PubMed ID12045096
JournalAnnu Rev Biochem
Year2002
Volume71
Pages221-46
AuthorsRees DC
TitleGreat metalloclusters in enzymology.
[81]
CommentsX-ray crystallography
PubMed ID12501184
JournalBiochemistry
Year2002
Volume41
Pages15557-65
AuthorsSchmid B, Einsle O, Chiu HJ, Willing A, Yoshida M, Howard JB, Rees DC
TitleBiochemical and structural characterization of the cross-linked complex of nitrogenase: comparison to the ADP-AlF4(-)-stabilized structure.
Related PDB1m1y,1m34
[82]
PubMed ID12401079
JournalInorg Chem
Year2002
Volume41
Pages5744-53
AuthorsLovell T, Torres RA, Han WG, Liu T, Case DA, Noodleman L
TitleMetal substitution in the active site of nitrogenase MFe(7)S(9) (M = Mo(4+), V(3+), Fe(3+)).
[83]
PubMed ID11782173
JournalJ Am Chem Soc
Year2002
Volume124
Pages216-24
AuthorsCoucouvanis D, Han J, Moon N
TitleSynthesis and characterization of sulfur-voided cubanes. Structural analogues for the MoFe(3)S(3) subunit in the nitrogenase cofactor.
[84]
CommentsX-ray crystallography
PubMed ID12133839
JournalJ Biol Chem
Year2002
Volume277
Pages35263-6
AuthorsMayer SM, Gormal CA, Smith BE, Lawson DM
TitleCrystallographic analysis of the MoFe protein of nitrogenase from a nifV mutant of Klebsiella pneumoniae identifies citrate as a ligand to the molybdenum of iron molybdenum cofactor (FeMoco).
Related PDB1h1l
[85]
PubMed ID12176981
JournalJ Biol Chem
Year2002
Volume277
Pages40106-11
AuthorsRangaraj P, Ludden PW
TitleAccumulation of 99Mo-containing iron-molybdenum cofactor precursors of nitrogenase on NifNE, NifH, and NifX of Azotobacter vinelandii.
[86]
PubMed ID11913144
JournalMet Ions Biol Syst
Year2002
Volume39
Pages75-119
AuthorsLawson DM, Smith BE
TitleMolybdenum nitrogenases: a crystallographic and mechanistic view.
[87]
CommentsX-RAY CRYSTALLOGRAPHY (1.16 ANGSTROMS)
Medline ID22204524
PubMed ID12215645
JournalScience
Year2002
Volume297
Pages1696-700
AuthorsEinsle O, Tezcan FA, Andrade SL, Schmid B, Yoshida M, Howard JB, Rees DC
TitleNitrogenase MoFe-protein at 1.16 A resolution: a central ligand in the FeMo-cofactor.
Related PDB1m1n
Related UniProtKBP07328,P07329
[88]
CommentsX-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS)
Medline ID21948294
PubMed ID11951047
JournalScience
Year2002
Volume296
Pages352-6
AuthorsSchmid B, Ribbe MW, Einsle O, Yoshida M, Thomas LM, Dean DR, Rees DC, Burgess BK
TitleStructure of a cofactor-deficient nitrogenase MoFe protein.
Related PDB1l5h
Related UniProtKBP07328,P07329
[89]
PubMed ID12506366
JournalChemistry
Year2003
Volume9
Pages76-87
AuthorsPickett CJ, Vincent KA, Ibrahim SK, Gormal CA, Smith BE, Best SP
TitleElectron-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.
[90]
PubMed ID15102840
JournalJ Biol Chem
Year2004
Volume279
Pages28276-82
AuthorsCorbett MC, Hu Y, Naderi F, Ribbe MW, Hedman B, Hodgson KO
TitleComparison of iron-molybdenum cofactor-deficient nitrogenase MoFe proteins by X-ray absorption spectroscopy: implications for P-cluster biosynthesis.
[91]
PubMed ID15549494
JournalJ Biol Inorg Chem
Year2004
Volume9
Pages1028-33
AuthorsJang SB, Jeong MS, Seefeldt LC, Peters JW
TitleStructural and biochemical implications of single amino acid substitutions in the nucleotide-dependent switch regions of the nitrogenase Fe protein from Azotobacter vinelandii.
Related PDB1xd8,1xd9,1xdb
[92]
PubMed ID14967020
JournalBiochemistry
Year2004
Volume43
Pages1787-97
AuthorsSen S, Igarashi R, Smith A, Johnson MK, Seefeldt LC, Peters JW
TitleA conformational mimic of the MgATP-bound "on state" of the nitrogenase iron protein.
Related PDB1rw4
[93]
PubMed ID15650336
JournalMol Cells
Year2004
Volume18
Pages374-82
AuthorsJeong MS, Jang SB
TitleStructural basis for the changes in redox potential in the nitrogenase Phe135Trp Fe protein with MgADP Bound.
Related PDB1xcp
[94]
PubMed ID16123301
JournalScience
Year2005
Volume309
Pages1377-80
AuthorsTezcan FA, Kaiser JT, Mustafi D, Walton MY, Howard JB, Rees DC
TitleNitrogenase complexes: multiple docking sites for a nucleotide switch protein.
Related PDB2afh,2afi,2afk
[95]
PubMed ID16616373
JournalJ Inorg Biochem
Year2006
Volume100
Pages1041-52
AuthorsSen S, Krishnakumar A, McClead J, Johnson MK, Seefeldt LC, Szilagyi RK, Peters JW
TitleInsights into the role of nucleotide-dependent conformational change in nitrogenase catalysis: Structural characterization of the nitrogenase Fe protein Leu127 deletion variant with bound MgATP.
Related PDB2c8v

comments
This enzyme is homologous to that from Klebsiella pneumoniae and Azotobacter vinelandii (M00114 in EzCatDB).
This enzyme, Nitrogenase, is composed of molybdenum-iron protein component (MoFe protein; component 1) and iron protein (Fe protein; component 2). The Fe protein component (component 2) transfer electrons to another enzyme component, MoFe protein (component 1).
The Fe protein component is a homodimer, which binds two MgATP molecules and one [4Fe-4S] cluster on the interface of the dimer.
The MoFe protein is a heterotetramer, composed of two alpha subunits and two beta subunits. The alpha subunit binds a cofactor called, FeMoco centre, which is composed of molybdenum, [7Fe-9S], and homocitrate, whereas the interface of the alpha and beta subunits binds a cofactor, called P cluster, that is [8Fe-7S].
This enzyme catalyzes the following reactions:
(A) Electron transfer from FMN of flavodoxin or [2Fe-2S] of ferredoxin to the [4Fe-4S] cluster of Fe protein:
(X) Binding of MgATP molecules to the Fe proteins induces the conformational change, leading to the reaction (B):
(B) Electron transfer from the [4Fe-4S] cluster to the P-cluster([8Fe-7S]):
(C) Hydrolysis of two ATP molecules to produce two ADP molecules, leading to the conformational change in disconnection between the Fe protein and MoFe protein, which inhibits the reaction (B) (Fe protein):
(D) Electron transfer from the P-cluster([8Fe-7S]) to the FeMoco centre:
(E) Hydrogenation (H2 production from 2 H+ ions) at the FeMoco centre:
(F) Nitorogenation (NH3 production from N2 and H+ ions) at the FeMoco centre:
###
This enzyme catalyzes N2 reduction:
N2 + 8 H+ + 8 e- => 2 NH3 + H2
(1) 2 H+ + 2 e- => H2
(2) N2 + 6 H+ + 6 e- => 2 NH3
Thus, the reactions (E) and (F) (or the MoFe protein cycle) consists of eight sequential one-electron steps [reactions, (A)-(D)], each of which corresponds to an Fe protein cycle (see [12], [86]).
###
(A) Electron transfer from FMN of flavodoxin or [2Fe-2S] of ferredoxin to the [4Fe-4S] cluster of Fe protein(see [93], [94]):
(B) Electron transfer from the [4Fe-4S] cluster to the P-cluster([8Fe-7S]):
(B1) Indirect transfer from the [4Fe-4S] cluster to the P-cluster through Cys94 (bound to [4Fe-4S]), Thr110 and Cys106 (bound to P-cluster).
(C) Hydrolysis of two ATP molecules to produce two ADP molecules on the Fe protein (see [16], [17], [18], [35], [91]):
(C1) The negative charge of gamma-phsohate group is stabilized by the magnesium ion bound to Ser15, and Lysine residues (Lys9, Lys14, Lys40), and mainchain amide group of Gly11.
(C2) Either Asp38 or Asp126 from the adjacent subunit acts as a general base to deprotonate and activate the water molecule. However, this water is not bound to the magnesium ion.
(C3) The activated water molecule makes a nucleophilic attack on the phosphorus atom of the gamma-phosphate.
(C4) The transition-state must be stabilized by the above stabilizer residues and cofactor.
(C5) The reaction completes.
(D) Electron transfer from the P-cluster([8Fe-7S]) to the FeMoco centre (see [20], [33], [74]):
(D1) Indirect transfer from the the P-cluster to the FeMoco centre through Cys53 (bound to P-cluster), Gln182 (bound to homocitrate), and homocitrate itself (bound to FeMoco).
(E) Hydrogenation (H2 production from 2 H+ ions) at the FeMoco centre (see [20], [73]):
(F) Nitorogenation (NH3 production from N2 and H+ ions) at the FeMoco centre (see [24], [87], [89]):

createdupdated
2005-04-252009-02-26


Copyright: Nozomi Nagano, JST & CBRC-AIST
Funded by PRESTO/Japan Science and Technology Corporation (JST) (December 2001 - November 2004)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2006)
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Funded by BIRD/Japan Science and Technology Corporation (JST) (September 2005 - September 2008)
Funded by BIRD/Japan Science and Technology Corporation (JST) (October 2007 - September 2010)
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