title: Molecular insights into nitrogenase FeMo cofactor insertion: the role of His 362 of the MoFe protein alpha subunit in FeMo cofactor incorporation.
abstract: The assembly of the complex iron-molybdenum cofactor (FeMoco) of nitrogenase molybdenum-iron (MoFe) protein has served as one of the central topics in the field of bioinorganic chemistry for decades. Here we examine the role of a MoFe protein residue (His alpha362) in FeMoco insertion, the final step of FeMoco biosynthesis where FeMoco is incorporated into its binding site in the MoFe protein. Our data from combined metal, activity and electron paramagnetic resonance analyses show that mutations of His alpha362 to small uncharged Ala or negatively charged Asp result in significantly reduced FeMoco accumulation in MoFe protein, indicating that His alpha362 plays a key role in the process of FeMoco insertion. Given the strategic location of His alpha362 at the entry point of the FeMoco insertion funnel, this residue may serve as one of the initial docking points for FeMoco insertion through transient ligand coordination and/or electrostatic interaction.
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 / Q000235:genetics; D004578:Electron Spin Resonance Spectroscopy; D004795:Enzyme Stability; D006639:Histidine / Q000235:genetics / Q000378:metabolism*; D050296:Microbial Viability; D008958:Models, Molecular; D008983:Molybdoferredoxin / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009591:Nitrogenase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D008983:Molybdoferredoxin; D021122:Protein Subunits; D006639:Histidine; D009591:Nitrogenase