Title page for ETD etd-11162006-104012


Type of Document Master's Thesis
Author Cotton, Marcia S.
Author's Email Address mcotto3@lsu.edu
URN etd-11162006-104012
Title MCD and Magnetization Studies of NifB/NifZ Minus Molybdenum Iron Protein from Azotobacter vinelandii
Degree Master of Science (M.S.)
Department Chemistry
Advisory Committee
Advisor Name Title
Brian J. Hales Committee Chair
Andrew W. Maverick Committee Member
George Stanley Committee Member
Keywords
  • vt mcd
  • magnetization
  • nifbnifz minus
  • mofe protein
Date of Defense 2006-11-10
Availability unrestricted
Abstract
This study reports a characterization of the iron-sulfur clusters of the MoFe protein expressed by the nifBnifZ deletion strain of A. vinelandii. VT-MCD spectroscopic and magnetization analysis of as-isolated ΔnifBΔnifZ MoFe protein were undertaken. VT-MCD spectra of as-isolated ΔnifBΔnifZ MoFe protein contains bands remarkably similar to those seen in as-isolated ΔnifH MoFe protein, assigned to arise from an [4Fe-4S]+ cluster. The low-temperature magnetization curve of as-isolated ΔnifBΔnifZ MoFe protein reveals the MCD spectra bands stem from an S = spin system with diamagnetic contributions of some other species. As-isolated ΔnifBΔnifZ MoFe protein is compared with as-isolated ΔnifH MoFe protein, which contains a P-cluster precursor, and the as-isolated ΔnifB MoFe protein, which contains a fully assembled P-cluster ([8Fe-7S)]. Using nonlinear regression, the experimental magnetization curve of as-isolated ΔnifBΔnifZ MoFe protein is compared and factored to contributions from as-isolated ΔnifH MoFe protein and as-isolated ΔnifB MoFe protein. The simulation suggests a 1:1 ratio of ΔnifH MoFe protein to ΔnifB MoFe protein. These results suggests the as-isolated ΔnifBΔnifZ MoFe protein has an αβ subunit pair that contains a P-cluster and the other αβ subunit pair contains a latent (defined as immature, or potential) P-cluster (two [4Fe-4S] or P-cluster precursor).
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