title: Identification of a nitrogenase FeMo cofactor precursor on NifEN complex.
abstract: The biosynthesis of the FeMo cofactor (FeMoco) of Azotobacter vinelandii nitrogenase presumably starts with the production of its Fe/S core by NifB (the nifB gene product). This core is subsequently processed on the alpha2beta2 tetrameric NifEN complex (formed by the nifE and nifN gene products). In this article, we identify a NifEN-bound FeMoco precursor form that can be converted to fully assembled FeMoco in a so-called FeMoco-maturation assay containing only purified components. We also establish that only molybdate, homocitrate, MgATP, and Fe protein are essential for FeMoco maturation. The FeMoco-maturation assay described here will further address the remaining questions related to the assembly mechanism of the ever-intriguing FeMoco.
authors.affiliation: Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697-3900, USA.
authors.forename: Yilin
authors.initials: Y
authors.lastname: Hu
authors.forename: Aaron W
authors.initials: AW
authors.lastname: Fay
authors.forename: Markus W
authors.initials: MW
authors.lastname: Ribbe
mesh_terms: D016948:Azotobacter vinelandii / Q000201:enzymology / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D008983:Molybdoferredoxin / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D008983:Molybdoferredoxin
references.citation: Biochemistry. 1997 Feb 11;36(6):1181-7
references.pmid: 9063865
references.citation: Proc Natl Acad Sci U S A. 2001 May 8;98(10):5521-5
references.pmid: 11331775
references.citation: Annu Rev Plant Physiol Plant Mol Biol. 2001 Jun;52:269-295
references.pmid: 11337399
references.citation: Biochemistry. 1998 Jul 21;37(29):10420-8
references.pmid: 9671511
references.citation: Appl Environ Microbiol. 1984 May;47(5):1027-30
references.pmid: 16346533
references.citation: Chem Rev. 1996 Nov 7;96(7):2983-3012
references.pmid: 11848849
references.citation: Science. 2002 Apr 12;296(5566):352-6
references.pmid: 11951047
references.citation: Science. 1992 Sep 18;257(5077):1653-9
references.pmid: 1529353
references.citation: J Biol Chem. 1998 Nov 6;273(45):29678-85
references.pmid: 9792679
references.citation: J Biol Chem. 1992 Oct 15;267(29):21179-86
references.pmid: 1400428
references.citation: J Biol Chem. 1998 Jul 3;273(27):16927-34
references.pmid: 9642255
references.citation: J Biol Chem. 2002 Apr 19;277(16):14299-305
references.pmid: 11823455
references.citation: Eur J Biochem. 1986 Oct 15;160(2):371-7
references.pmid: 3533537
references.citation: J Biol Chem. 2000 Jun 9;275(23):17631-8
references.pmid: 10837496
references.citation: Biochemistry. 2004 Feb 17;43(6):1401-9
references.pmid: 14769015
references.citation: J Biol Chem. 1999 Jul 9;274(28):19778-84
references.pmid: 10391920
references.citation: J Biol Chem. 2001 May 11;276(19):15968-74
references.pmid: 11279153
references.citation: J Biol Chem. 1995 Mar 3;270(9):4432-7
references.pmid: 7876209
references.citation: Chem Rev. 1996 Nov 7;96(7):2965-2982
references.pmid: 11848848
references.citation: Anal Biochem. 1967 Aug;20(2):325-34
references.pmid: 4293646
references.citation: Proc Natl Acad Sci U S A. 1986 Mar;83(6):1636-40
references.pmid: 3006060
references.citation: J Biol Chem. 2004 Jul 2;279(27):28276-82
references.pmid: 15102840
references.citation: J Biol Chem. 1995 Nov 10;270(45):26890-6
references.pmid: 7592933
references.citation: J Biol Chem. 2004 Dec 24;279(52):54963-71
references.pmid: 15485884
references.citation: J Biol Chem. 1995 Oct 20;270(42):24745-52
references.pmid: 7559591
references.citation: Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11250-5
references.pmid: 9326595
references.citation: J Bacteriol. 1987 Apr;169(4):1784-6
references.pmid: 3470286
references.citation: J Bacteriol. 1999 May;181(9):2797-801
references.pmid: 10217770
references.citation: Biochemistry. 1998 Sep 8;37(36):12611-23
references.pmid: 9730834
references.citation: J Biol Chem. 1994 Jan 14;269(2):1154-8
references.pmid: 8288575
references.citation: Biochim Biophys Acta. 1980 Jul 10;614(1):196-209
references.pmid: 6930977
references.citation: J Biol Chem. 1996 Feb 23;271(8):4256-60
references.pmid: 8626771
references.citation: Chem Rev. 2004 Feb;104(2):1159-73
references.pmid: 14871152
references.citation: J Bacteriol. 1987 Apr;169(4):1547-53
references.pmid: 3470285
references.citation: Proc Natl Acad Sci U S A. 1989 Aug;86(16):6082-6
references.pmid: 2668954
references.citation: Science. 2002 Sep 6;297(5587):1696-700
references.pmid: 12215645
references.citation: J Biol Chem. 2002 Jun 28;277(26):23469-76
references.pmid: 11978793
references.citation: Curr Opin Chem Biol. 2000 Oct;4(5):559-66
references.pmid: 11006545
references.citation: J Bacteriol. 1994 May;176(9):2747-50
references.pmid: 8169227
references.citation: J Biol Chem. 1989 Jun 15;264(17):10088-95
references.pmid: 2785995
references.citation: J Biol Chem. 1985 Apr 10;260(7):3891-4
references.pmid: 3856566
references.citation: Crit Rev Biochem Mol Biol. 2003;38(4):351-84
references.pmid: 14551236
references.citation: J Biol Chem. 2002 Oct 18;277(42):40106-11
references.pmid: 12176981
references.citation: J Biol Chem. 1987 Oct 15;262(29):14327-32
references.pmid: 3477546
references.citation: J Bacteriol. 1991 Sep;173(17):5403-5
references.pmid: 1885520