Oxidation-reduction potential dependence of photosystem II carbonic anhydrase in maize thylakoids.
In characterizing the carbonic anhydrase (CA) found in maize thylakoid membranes, it was observed that the enzyme's activity was inhibited somewhat when the Hill oxidant, ferricyanide, was given in the dark [Stemler, A. (1986) Biochim. Biophys. Acta 850, 97-107]. In the present work, a redox titration of this effect shows that the CA activity is mediated by a component that has a midpoint potential (Em) of about 485 mV at pH 6.5 and a pH dependence of 60 mV/pH. These redox titration characteristics are identical to those of the redox mediator "D480", which modulates formate and bicarbonate binding affinity to photosystem II (PS II). Bicarbonate binds to PS II more readily, and CA activity is higher, when D480 is reduced, whereas both bicarbonate binding and thylakoid-bound CA activity are low when D480 is oxidized in the dark by ferricyanide. Both the low bicarbonate binding affinity and the low CA activity induced by the presence of ferricyanide are reversed by a single saturating flash of light. In contrast, the activity of soluble CA, which is extracted from maize mesophyll cytosol, does not exhibit any redox dependence in the range 400-550 mV. Furthermore, thylakoid-bound CA activity is inhibited by 5 mM ZnCl2 by as much as 75%, whereas the activity of soluble CA shows no significant decrease induced by ZnCl2. Also, at a medium potential of 400 mV, ferricyanide (1 mM) inhibits soluble CA activity by 88% and thylakoid-bound CA activity by only 18%. It is concluded from these results that CA activity observed in thylakoids arises from CA inherent to PS II and is not some form of contamination by soluble CA. Possible roles of CA in PS II reaction mechanisms are discussed.
Moubarak-Milad
M
M
Section of Plant Biology, University of California, Davis 95616.
Stemler
A
A
D001639:Bicarbonates / Q000378:metabolism; D002256:Carbonic Anhydrases / Q000737:chemistry* / Q000378:metabolism; D002734:Chlorophyll / Q000378:metabolism; D007425:Intracellular Membranes / Q000201:enzymology* / Q000378:metabolism; D045342:Light-Harvesting Protein Complexes; D010084:Oxidation-Reduction; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000378:metabolism; D045332:Photosystem II Protein Complex; D012995:Solubility; D003313:Zea mays / Q000201:enzymology*
D001639:Bicarbonates; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D002734:Chlorophyll; D002256:Carbonic Anhydrases