Molecular insights into nitrogenase FeMoco insertion--the role of His 274 and His 451 of MoFe protein alpha subunit.
The final step of FeMo cofactor (FeMoco) assembly involves the insertion of FeMoco into its binding site in the molybdenum-iron (MoFe) protein of nitrogenase. Here we examine the role of His alpha274 and His alpha451 of Azotobacter vinelandii MoFe protein in this process. Our results from combined metal, activity, EPR, stability and insertion analyses show that mutations of His alpha274 and/or His alpha451, two of the histidines that belong to a so-called His triad, to small uncharged Ala specifically reduce the accumulation of FeMoco in MoFe protein. This observation indicates that the enrichment of histidines at the His triad is important for FeMoco insertion and that the His triad potentially serves as an intermediate docking point for FeMoco through transitory ligand coordination and/or electrostatic interaction.
Fay
Aaron W
AW
Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697-3900, USA.
Hu
Yilin
Y
Schmid
Benedikt
B
Ribbe
Markus W
MW
D016948:Azotobacter vinelandii / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D006639:Histidine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D008983:Molybdoferredoxin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D009591:Nitrogenase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
D008983:Molybdoferredoxin; D021122:Protein Subunits; D006639:Histidine; D009591:Nitrogenase