Cysteine dioxygenase structures from pH 4 to 9: consistent Cys-persulfenate formation at intermediate pH and a Cys-bound enzyme at higher pH Public Deposited

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  • Mammalian cysteine dioxygenase (CDO) is a mononuclear non-heme iron protein that catalyzes 2 the conversion of cysteine (Cys) to cysteine sulfinic acid (CSA) by an unclarified mechanism. 3 One structural study revealed a Cys-persulfenate (or Cys-persulfenic acid) formed in the active 4 site, but quantum mechanical calculations have been used to support arguments that it is not an 5 energetically feasible reaction intermediate. Here, we report a series of high-resolution structures 6 of CDO soaked with Cys at pH values from 4 to 9. Cys binding is minimal at pH≤5 and 7 persulfenate formation is consistently seen at pH values between 5.5 and 7. Also, a structure 8 determined using laboratory-based X-ray diffraction shows that the persulfenate, with an 9 apparent average O-O separation distance of ~1.8 Å is not an artifact of synchrotron radiation. At 10 pH≥8, the active site iron shifts from 4- to 5-coordinate, and Cys soaks reveal a complex with 11 Cys, but no dioxygen, bound. This ‘Cys-only’ complex differs in detail from a previously 12 published ‘Cys-only’ complex which we reevaluate and conclude is not reliable. The high-13 resolution structures presented here do not resolve the CDO mechanism, but do imply that an 14 iron-bound persulfenate (or persulfenic acid) is energetically accessible in the CDO active site, 15 and that CDO active site chemistry in the crystals is influenced by protonation/deprotonation 16 events with effective pKa values near ~5.5 and ~7.5 that influence Cys binding and oxygen 17 binding/reactivity, respectively. Furthermore, this work provides reliable ligand-bound models 18 for guiding future mechanistic considerations.
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  • Driggers, C.M., Cooley, R.B., Sankaran, B., Hirschberger, L.L, Stipanuk, M.H., & Karplus, P.A. (2013). Cysteine dioxygenase structures from pH 4 to 9: Consistent cys-persulfenate formation at intermediate pH and a cys-bound enzyme at higher pH. Journal of Molecular Biology, 425(17), 3121-3136. doi:10.1016/j.jmb.2013.05.028
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  • description.provenance : Approved for entry into archive by Deanne Bruner(deanne.bruner@oregonstate.edu) on 2013-10-04T23:27:40Z (GMT) No. of bitstreams: 2KarplusPAndrewBiochemistryBiophysicsCysteineDioxygenaseStructures.pdf: 1335219 bytes, checksum: 7cfe1b9bafc26409d6da115bb52d5b01 (MD5)KarplusPAndrewBiochemistryBiophysicsCysteineDioxygenaseStructures(SupplementaryInformation).pdf: 309385 bytes, checksum: 1e07aee150de201501b1cb3559708c94 (MD5)
  • description.provenance : Made available in DSpace on 2013-10-04T23:27:40Z (GMT). No. of bitstreams: 2KarplusPAndrewBiochemistryBiophysicsCysteineDioxygenaseStructures.pdf: 1335219 bytes, checksum: 7cfe1b9bafc26409d6da115bb52d5b01 (MD5)KarplusPAndrewBiochemistryBiophysicsCysteineDioxygenaseStructures(SupplementaryInformation).pdf: 309385 bytes, checksum: 1e07aee150de201501b1cb3559708c94 (MD5) Previous issue date: 2013-09-09
  • This is an author's peer-reviewed final manuscript, as accepted by the publisher.
  • description.provenance : Submitted by Deanne Bruner (deanne.bruner@oregonstate.edu) on 2013-10-04T23:27:03ZNo. of bitstreams: 2KarplusPAndrewBiochemistryBiophysicsCysteineDioxygenaseStructures.pdf: 1335219 bytes, checksum: 7cfe1b9bafc26409d6da115bb52d5b01 (MD5)KarplusPAndrewBiochemistryBiophysicsCysteineDioxygenaseStructures(SupplementaryInformation).pdf: 309385 bytes, checksum: 1e07aee150de201501b1cb3559708c94 (MD5)

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