Characterization of a variant iron protein of nitrogenase that is impaired in its ability to adopt the MgATP-induced conformational change.
Abstract
An Azotobacter vinelandii nitrogenase iron protein mutant has been created which contains an alanine to glycine substitution at amino acid 157. The strain expressing this mutant Fe protein is able to grow under nitrogen-fixing conditions. This contrasts with an A. vinelandii strain described previously which is unable to grow under nitrogen-fixing conditions and which expresses an Fe protein variant that has an alanine to serine mutation at position 157. The A157S Fe protein was unable to support substrate reduction by nitrogenase because of an inability to undergo a required MgATP-induced conformational change. Although the A157G strain grows at 55% of the rate of the wild-type strain, purified A157G Fe protein is only able to support substrate reduction in in vitro assays at a rate that is approximately 20% of the rate supported by the wild-type Fe protein. Electron paramagnetic resonance, circular dichroism spectroscopies, and enzymatic activity data indicate that the A157G Fe protein adopts the correct conformation upon the binding of MgATP. However, kinetic studies using chelation show that this protein undergoes the conformational change more slowly than the wild-type protein. Thus, this mutant has lower activity because ...Continue Reading
References
Citations
NifEN-B complex of Azotobacter vinelandii is fully functional in nitrogenase FeMo cofactor assembly.
The chaperone GroEL is required for the final assembly of the molybdenum-iron protein of nitrogenase
Related Concepts
Related Feeds
ASBMB Publications
The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.