Biochemical analysis of the recombinant Fur (ferric uptake regulator) protein from Anabaena PCC 7119: factors affecting its oligomerization state.
作者： José A. HERNÁNDEZ , M. Teresa BES , María F. FILLAT , José L. NEIRA , M. Luisa PELEATO
DOI: 10.1042/BJ20020135
关键词: Electrospray ionization 、 DTNB 、 Anabaena 、 Cysteine 、 Polyclonal antibodies 、 Biology 、 Recombinant DNA 、 Matrix-assisted laser desorption/ionization 、 Biochemistry 、 Polyacrylamide gel electrophoresis
摘要: Fur (ferric uptake regulator) protein is a DNA-binding which regulates iron-responsive genes. Recombinant from the nitrogen-fixing cyanobacterium Anabaena PCC 7119 has been purified and characterized, polyclonal antibodies obtained. The experimental data show that dimerizes in solution with involvement of disulphide bridges. Cross-linking experiments MALDI-TOF (matrix-assisted laser desorption/ionization time flight) MS also several oligomerization states Fur, equilibrium these forms depends on concentration ionic strength. In intact recombinant four cysteine residues out five were inert towards DTNB [5,5'-dithiobis-(2-nitrobenzoic acid)], their modification required sodium borohydride. Metal analysis electrospray ionization revealed neither zinc nor other metals are present this protein. Purified bound to its own promoter gel-shift assays. was shown be constitutive cells, no significant difference expression cells grown under iron-sufficient compared iron-deficient conditions.
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