A carboxylic residue at the high-affinity, Mn-binding site participates in the binding of iron cations that block the site.
The role of carboxylic residues at the high-affinity, Mn-binding site in the ligation of iron cations blocking the site [Biochemistry 41 (2000) 5854] was studied, using a method developed to extract the iron cations blocking the site. We found that specifically bound Fe(III) cations can be extracted with citrate buffer at pH 3.0. Furthermore, citrate can also prevent the photooxidation of Fe(II) cations by YZ. Participation of a COOH group(s) in the ligation of Fe(III) at the high-affinity site was investigated using 1-ethyl-3-[(3-dimethylamino)propyl] carbodiimide (EDC), a chemical modifier of carboxylic amino acid residues. Modification of the COOH groups inhibits the light-induced oxidation of exogenous Mn(II) cations by Mn-depleted photosystem II (PSII[-Mn]) membranes. The rate of Mn(II) oxidation saturates at > or = 10 microM in PSII(-Mn) membranes and > or = 500 microM in EDC-treated PSII (-Mn) samples. Intact PSII(-Mn) membranes have only one site for Mn(II) oxidation via YZ (dissociation constant, Kd = 0.64 microM), while EDC-treated PSII(-Mn) samples have two sites (Kd = 1.52 and 22 microM; the latter is the low-affinity site). When PSII(-Mn) membranes were incubated with Fe(II) before modifier treatment (to block the high-affinity site) and the blocking iron cations were extracted with citrate (pH 3.0) after modification, the membranes contained only one site (Kd = 2.3 microM) for exogenous Mn(II) oxidation by Y(Z)() radical. In this case, the rate of electron donation via YZ saturated at a Mn(II) concentration > or = 15 microM. These results indicate that the carboxylic residue participating in Mn(II) coordination and the binding of oxidized manganese cations at the HAZ site is protected from the action of the modifier by the iron cations blocking the HAZ site. We concluded that the carboxylic residue (D1 Asp-170) participating in the coordination of the manganese cation at the HAZ site (Mn4 in the tetranuclear manganese cluster [Science 303 (2004) 1831]) is also involved in the ligation of the Fe cation(s) blocking the high-affinity Mn-binding site.
Basic Sciences Center, National Renewable Energy Laboratory, Golden, CO 80401, USA.
Boris K
BK
Semin
Michael
M
Seibert
D001665:Binding Sites; D002412:Cations / Q000737:chemistry; D004579:Electron Transport; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000494:pharmacology; D007501:Iron / Q000737:chemistry* / Q000378:metabolism*; D008345:Manganese / Q000737:chemistry* / Q000378:metabolism*; D045332:Photosystem II Protein Complex / Q000378:metabolism; D010936:Plant Extracts / Q000378:metabolism; D018724:Spinacia oleracea / Q000187:drug effects / Q000201:enzymology
D002412:Cations; D045332:Photosystem II Protein Complex; D010936:Plant Extracts; D008345:Manganese; D007501:Iron; D005022:Ethyldimethylaminopropyl Carbodiimide