title: Effects of ethylene glycol and methanol on ammonia-induced structural changes of the oxygen-evolving complex in photosystem II.
abstract: Ammonia is an inhibitor of water oxidation and a structural analogue for substrate water, making it a valuable probe for the structural properties of the possible substrate-binding site on the oxygen-evolving complex (OEC) in photosystem II (PSII). By using the NH(3)-induced upshift of the 1365 cm(-)(1) IR mode in the S(2)Q(A)(-)/S(1)Q(A) spectrum and the NH(3)-modified S(2) state EPR signals of PSII as spectral probes, we found that ethylene glycol has clear effects on the binding properties of the NH(3)-specific site on the OEC. Our results show that in PSII samples containing 30% (v/v) ethylene glycol, the affinity of the NH(3)-specific binding site on the OEC is estimated to be more than 10 times lower than that in PSII samples containing 0.4 M sucrose. In addition, our results show that the NH(3)-induced upshift of the 1365 cm(-)(1) IR mode in the S(2)Q(A)(-)/S(1)Q(A) spectrum is dependent on the concentration of ethylene glycol, but not dependent on the concentration of sucrose (up to 1.5 M) or methanol (up to 5.4 M). By comparing the concentration dependence of sucrose and ethylene glycol on NH(3)-induced spectral change and also by comparing the sucrose and ethylene glycol data at similar concentrations ( approximately 1 M), we conclude that ethylene glycol has a clear effect on the NH(3)-induced spectral changes. Furthermore, our results also show that ethylene glycol alters the steric requirement of the amine effect on the upshift of the 1365 cm(-)(1) mode in the S(2)Q(A)(-)/S(1)Q(A) spectrum. In PSII samples containing 30% (v/v) ethylene glycol, only NH(3), not other bulkier amines (e.g., Tris, AEPD, and CH(3)NH(2)), has a clear effect on the upshift of the 1365 cm(-)(1) mode in the S(2)Q(A)(-)/S(1)Q(A) spectrum; in contrast, in PSII samples containing 0.4 M sucrose, both NH(3) and CH(3)NH(2) have a clear effect. On the basis of the results mentioned above, we propose that ethylene glycol acts directly or indirectly to decrease the affinity or limit the accessibility of NH(3) and CH(3)NH(2) to the NH(3)-specific binding site on the OEC in PSII. Finally, we also applied the same approach to test whether methanol is able to compete with ammonia on its binding site on the OEC. We found that 4% (v/v) methanol does not have any significant effect on the NH(3)-induced upshift of the 1365 cm(-)(1) mode in the S(2)Q(A)(-)/S(1)Q(A) spectrum and the NH(3)-modified S(2) state g = 2 multiline EPR signal. Our results suggest that methanol is unable to compete with NH(3) upon binding to the Mn site of the OEC that gives rise to the altered S(2) state g = 2 multiline EPR signal.
authors: Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan 11529, Republic of China.
authors: Cheng-Hao
authors: CH
authors: Fang
authors: Kuo-An
authors: KA
authors: Chiang
authors: Chung-Hsien
authors: CH
authors: Hung
authors: Keejong
authors: K
authors: Chang
authors: Shyue-Chu
authors: SC
authors: Ke
authors: Hsiu-An
authors: HA
authors: Chu
mesh_terms: D000641:Ammonia / Q000737:chemistry* / Q000378:metabolism; D000643:Ammonium Chloride / Q000737:chemistry; D016227:Benzoquinones / Q000737:chemistry; D001667:Binding, Competitive; D004578:Electron Spin Resonance Spectroscopy; D019855:Ethylene Glycol / Q000737:chemistry*; D008345:Manganese / Q000378:metabolism; D000432:Methanol / Q000737:chemistry* / Q000378:metabolism; D010100:Oxygen / Q000737:chemistry* / Q000378:metabolism; D045332:Photosystem II Protein Complex / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D017550:Spectroscopy, Fourier Transform Infrared; D018724:Spinacia oleracea; D013237:Stereoisomerism
chemical_list: D016227:Benzoquinones; D045332:Photosystem II Protein Complex; D000643:Ammonium Chloride; C025232:semiquinone radicals; D008345:Manganese; D000641:Ammonia; D019855:Ethylene Glycol; D010100:Oxygen; D000432:Methanol