Redox-dependent conformational selection in a Cys4Fe2S2 ferredoxin

Biochemistry
Thomas C PochapskyR Pejchal

Abstract

Putidaredoxin (Pdx), a Cys4Fe2S2 ferredoxin from Pseudomonas putida, exhibits redox-dependent binding to its physiological redox partner, cytochrome P450(cam) (CYP101), with the reduced form of Pdx (Pdx(r)) binding with greater affinity to oxidized camphor-bound CYP101 than the oxidized form, Pdx(o). It has been previously shown that Pdx(o) is more dynamic than Pdx(r) on all accessible time scales, and it has been proposed that Pdx(r) samples only a fraction of the conformational substates populated by Pdx(o) on a time average. It is postulated that the ensemble subset populated by Pdx(r) is the same subset that binds CYP101, providing a mechanism for coupling the Pdx oxidation state to binding affinity for CYP101. Evidence from a variety of sources, including redox-dependent shifts of 15N and 13C resonances, indicates that the metal cluster binding loop of Pdx is the primary determinant of redox-dependent conformational selection. Patterns of paramagnetic effects suggest that the metal cluster binding loop contracts around the metal cluster upon reduction, possibly due to the strengthening of hydrogen bonds between the sulfur atoms of the metal cluster and the surrounding polypeptide NH and OH groups. Effects of this perturbat...Continue Reading

References

Feb 5, 1999·Journal of Molecular Biology·D W Miller, D A Agard
Jan 15, 2000·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Michael J Sutcliffe, Nigel S Scrutton

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Citations

Jul 16, 2008·Analytical and Bioanalytical Chemistry·Emre M Isin, F Peter Guengerich
Apr 4, 2012·Biochemistry·Eliana K AsciuttoThomas C Pochapsky
Apr 16, 2009·Biochemistry·Susan Sondej PochapskyThomas C Pochapsky
Jan 26, 2010·The Journal of Biological Chemistry·Jed N LampePaul R Ortiz de Montellano
Aug 6, 2013·The Journal of Physical Chemistry. B·Ravinder Kaur, Kara L Bren
May 26, 2004·Biochimica Et Biophysica Acta·Vytas ReipaVincent L Vilker
Jun 3, 2008·Structure·Bo OuYangThomas C Pochapsky
Apr 30, 2019·Accounts of Chemical Research·Thomas C Pochapsky, Susan Sondej Pochapsky

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