title: Calcium controls the assembly of the photosynthetic water-oxidizing complex: a cadmium(II) inorganic mutant of the Mn4Ca core.
abstract: Perturbation of the catalytic inorganic core (Mn4Ca1OxCly) of the photosystem II-water-oxidizing complex (PSII-WOC) isolated from spinach is examined by substitution of Ca2+ with cadmium(II) during core assembly. Cd2+ inhibits the yield of reconstitution of O2-evolution activity, called photoactivation, starting from the free inorganic cofactors and the cofactor-depleted apo-WOC-PSII complex. Ca2+ affinity increases following photooxidation of the first Mn2+ to Mn3+ bound to the 'high-affinity' site. Ca2+ binding occurs in the dark and is the slowest overall step of photoactivation (IM1-->IM1* step). Cd2+ competitively blocks the binding of Ca2+ to its functional site with 10- to 30-fold higher affinity, but does not influence the binding of Mn2+ to its high-affinity site. By contrast, even 10-fold higher concentrations of Cd2+ have no effect on O2-evolution activity in intact PSII-WOC. Paradoxically, Cd2+ both inhibits photoactivation yield, while accelerating the rate of photoassembly of active centres 10-fold relative to Ca2+. Cd2+ increases the kinetic stability of the photooxidized Mn3+ assembly intermediate(s) by twofold (mean lifetime for dark decay). The rate data provide evidence that Cd2+ binding following photooxidation of the first Mn3+, IM1-->IM1*, causes three outcomes: (i) a longer intermediate lifetime that slows IM1 decay to IM0 by charge recombination, (ii) 10-fold higher probability of attaining the degrees of freedom (either or both cofactor and protein d.f.) needed to bind and photooxidize the remaining 3 Mn2+ that form the functional cluster, and (iii) increased lability of Cd2+ following Mn4 cluster assembly results in (re)exchange of Cd2+ by Ca2+ which restores active O2-evolving centres. Prior EPR spectroscopic data provide evidence for an oxo-bridged assembly intermediate, Mn3+(mu-O2(-))Ca2+, for IM1*. We postulate an analogous inhibited intermediate with Cd2+ replacing Ca2+.
authors.affiliation: Department of Chemistry, Princeton University, Princeton, NJ 08544, USA.
authors.forename: John E
authors.initials: JE
authors.lastname: Bartlett
authors.forename: Sergei V
authors.initials: SV
authors.lastname: Baranov
authors.forename: Gennady M
authors.initials: GM
authors.lastname: Ananyev
authors.forename: G Charles
authors.initials: GC
authors.lastname: Dismukes
mesh_terms: D002104:Cadmium / Q000737:chemistry*; D002118:Calcium / Q000737:chemistry*; D007700:Kinetics; D008345:Manganese / Q000737:chemistry*; D008956:Models, Chemical; D010777:Photochemistry; D045332:Photosystem II Protein Complex / Q000737:chemistry*; D018724:Spinacia oleracea / Q000737:chemistry; D014867:Water / Q000737:chemistry*
chemical_list: D045332:Photosystem II Protein Complex; D002104:Cadmium; D014867:Water; D008345:Manganese; D002118:Calcium