title: pH-induced conformational change of IscU at low pH correlates with protonation/deprotonation of two conserved histidine residues.
abstract: IscU, the scaffold protein for the major iron-sulfur cluster biosynthesis pathway in microorganisms and mitochondria (ISC pathway), plays important roles in the formation of [2Fe-2S] and [4Fe-4S] clusters and their delivery to acceptor apo-proteins. Our laboratory has shown that IscU populates two distinct, functionally relevant conformational states, a more structured state (S) and a more dynamic state (D), that differ by cis/trans isomerizations about two peptidyl-prolyl peptide bonds [Kim, J. H., Tonelli, M., and Markley, J. L. (2012) Proc. Natl. Acad. Sci. U.S.A., 109, 454-459. Dai Z., Tonelli, M., and Markley, J. L. (2012) Biochemistry, 51, 9595-9602. Cai, K., Frederick, R. O., Kim, J. H., Reinen, N. M., Tonelli, M., and Markley, J. L. (2013) J. Biol. Chem., 288, 28755-28770]. Here, we report our findings on the pH dependence of the D ⇄ S equilibrium for Escherichia coli IscU in which the D-state is stabilized at low and high pH values. We show that the lower limb of the pH dependence curve results from differences in the pKa values of two conserved histidine residues (His10 and His105) in the two states. The net proton affinity of His10 is about 50 times higher and that of His105 is 13 times higher in the D-state than in the S-state. The origin of the high limb of the D ⇄ S pH dependence remains to be determined. These results show that changes in proton inventory need to be taken into account in the steps in iron-sulfur cluster assembly and transfer that involve transitions of IscU between its S- and D-states.
authors: Biophysics Graduate Program, ‡Biochemistry Department, and §National Magnetic Resonance Facility at Madison, Biochemistry Department, University of Wisconsin-Madison , Madison, Wisconsin 53715, United States.
authors: Ziqi
authors: Z
authors: Dai
authors: Jin Hae
authors: JH
authors: Kim
authors: Marco
authors: M
authors: Tonelli
authors: Ibrahim K
authors: IK
authors: Ali
authors: John L
authors: JL
authors: Markley
mesh_terms: D000595:Amino Acid Sequence; D017124:Conserved Sequence; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D006639:Histidine / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007506:Iron-Sulfur Proteins / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: D029968:Escherichia coli Proteins; D007506:Iron-Sulfur Proteins; C411665:IscU protein, E coli; D006639:Histidine