Formation of radicals from singlet oxygen produced during photoinhibition of isolated light-harvesting proteins of photosystem II

• Published in: Biochimica et biophysica acta 2004-01, Vol.1608 (1), p.63-73
• Main Authors: Rinalducci, Sara, Pedersen, Jens Z., Zolla, Lello
• Format: Article
• Language: English
• Subjects: Electron Spin Resonance Spectroscopy; Electrophoresis, Polyacrylamide Gel; ESR; Light; Light-harvesting complex; Light-Harvesting Protein Complexes - chemistry; Light-Harvesting Protein Complexes - isolation & purification; Oxygen; Photochemistry; Photoinhibition; photosynthesis; Photosystem II Protein Complex - chemistry; Protein degradation; Reactive oxygen species; Singlet oxygen; Singlet Oxygen - analysis; Singlet Oxygen - chemistry; Spin Trapping; Spinacia oleracea; Time Factors
• ISSN: 0005-2728; 0006-3002; 1879-2650
• DOI: 10.1016/j.bbabio.2003.10.009

Electron spin resonance spectroscopy and liquid chromatography have been used to detect radical formation and fragmentation of polypeptides during photoinhibition of purified major antenna proteins, free of protease contaminants. In the absence of oxygen and light, no radicals were observed and there was no damage to the proteins. Similarly illumination of the apoproteins did not induce any polypeptide fragmentation, suggesting that chlorophyll, light and atmospheric oxygen are all participating in antenna degradation. The use of TEMP and DMPO as spin traps showed that protein damage initiates with generation of 1O 2, presumably from a triplet chlorophyll, acting as a Type II photosensitizer which attacks directly the amino acids causing a complete degradation of protein into small fragments, without the contribution of proteases. Through the use of scavengers, it was shown that superoxide and H 2O 2 were not involved initially in the reaction mechanism. A higher production of radicals was observed in trimers than in monomeric antenna, while radical production is strongly reduced when antennae were organized in the photosystem II (PSII) complex. Thus, monomerization of antennae as well as their incorporation into the PSII complex seem to represent physiologically protected forms. A comparison is made of the photoinhibition mechanisms of different photosynthetic systems.