Elucidation of a [4Fe-4S] cluster degradation pathway: rapid kinetic studies of the degradation of Chromatium vinosum HiPIP

Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry
M W FosterJ A Cowan

Abstract

Irreversible disassembly of the 4Fe-4S cluster in Chromatium vinosum high-potential iron protein (HiPIP) has been investigated in the presence of a low concentration of guanidinium hydrochloride. From the dependence of degradation rate on [H+], it is deduced that at least three protons are required to trigger efficient cluster degradation. Under these conditions the protonated cluster shows broadened Mössbauer signals, but delta EQ (1.1 mm/s) and delta (0.44 mm/s) are similar to the native form. Collapse of the protonated transition state complex, revealed by rapid-quench Mössbauer experiments, occurs with a measured rate constant kobs approximately 0.72 +/- 0.35 s-1 that is consistent with results from time-resolved electronic absorption and fluorescence (kobs approximately 0.4 +/- 0.1 s-1) and EPR (kobs approximately 0.62 +/- 0.18 s-1) measurements. Apparently, guanidinium hydrochloride serves to perturb the tertiary structure of the protein, facilitating protonation of the cluster, but not degradation per se. Release of iron ions occurs even more slowly with kobs approximately 0.07 +/- 0.02 s-1, as determined by the appearance of the g = 4.3 EPR signal. Proton-mediated cluster degradation is sensitive to the oxidation state ...Continue Reading

Citations

Dec 4, 2004·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Sónia S LealCláudio M Gomes
Apr 17, 2002·Accounts of Chemical Research·Pernilla Wittung-Stafshede
Jun 16, 2004·Archives of Biochemistry and Biophysics·Catherine L Higgins, Pernilla Wittung-Stafshede

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