MOSSBAUER, ELECTRON-PARAMAGNETIC-RES, AND MAGNETIZATION STUDIES OF THE AZOTOBACTER-VINELANDII FE PROTEIN - EVIDENCE FOR A [4FE-4S]1+ CLUSTER WITH SPIN S=3/2

  • 1 January 1985
    • journal article
    • research article
    • Vol. 260  (20) , 1160-1173
Abstract
We have studied the Fe protein (Av2) of the Azotobacter vinelandii nitrogenase system with Mossbauer and EPR spectroscopies and magnetic susceptometry. In the oxidized state the protein exhibits Mossbauer spectra typical of diamagnetic [4Fe-4S]2+ clusters. Addition of Mg .cntdot. ATP or Mg .cntdot. ADP causes a pronounced decline in the quadrupole splitting of the Moessbauer spectra of the oxidized protein. Our studies show that reduced Av2 in the native state is heterogeneous. Approximately half of the molecules contain a [4Fe-4S]1+ cluster with electronic spin S = 1/2 and half contain a [4Fe-4S]1+ cluster with spin S = 3/2. The former yields the characteristic g = 1.94 EPR signal whereas the latter exhibits signals around g = 5. The magnetization of reduced Av2 is dominated by the spin S = 3/2 form of its [4Fe-4S]1+ clusters. These results explain a long standing puzzle, namely why the integrated spin intensity of the g = 1.94 EPR signal is substantially less than 1 spin/4 Fe atoms. In 50% ethylene glycol, 90% of the clusters are in the spin S = 1/2 form whereas, in 0.4 M urea, 85% are in the S = 3/2 form. In 0.4 M urea, the EPR spectrum of reduced Av2 exhibits well defined resonances at g = 5.8 and 515, which we assign to the S = 3/2 system. The EPR and Moessbauer studies yield a zero-field splitting of 2D .apprxeq. -5 cm-1 for this S = 3/2 state.