Photosynthetica 2006, 44(1):116-124 | DOI: 10.1007/s11099-005-0166-z

Radiation inactivation analysis of thylakoid protein kinase systems in light and in darkness

S. C. Lee1, L. F. Chien2,*, R. C. Van1, Y. Y. Hsiao1, J. L. Hong1, R. L. Pan1
1 Department of Life Sciences, College of Life Sciences, National Tsing Hua University, Hsin Chu, Taiwan
2 Department of Life Sciences, College of Life Sciences, National Chung Hsing University, Tai Chung, Taiwan

Chloroplast thylakoid contains several membrane-bound protein kinases that phosphorylate thylakoid polypeptides for the regulation of photosynthesis. Thylakoid protein phosphorylation is activated when the plastoquinone pool is reduced either by light-dependent electron flow through photosystem 2 (PS2) or by adding exogenous reductants such as durohydroquinone in the dark. The major phosphorylated proteins on thylakoid are components of light-harvesting complex 2 (LHC2) and a PS2 associated 9 kDa phosphoprotein. Radiation inactivation technique was employed to determine the functional masses of various kinases for protein phosphorylation in thylakoids. Under the photosynthetically active radiation (PAR), the apparent functional masses of thylakoid protein kinase systems (TPKXs) for catalyzing phosphorylation of LHC2 27 and 25 kDa polypeptides were 540±50 and 454±35 kDa as well as it was 448±23 kDa for PS2 9 kDa protein phosphorylation. Furthermore, the functional sizes of dark-regulated TPKXs for 25 and 9 kDa proteins were 318±25 and 160±8 kDa. The 9 kDa protein phosphorylation was independent of LHC2 polypeptides phosphorylation with regard to its TPKX functional mass. Target size analysis of protein phosphorylation mentioned above indicates that thylakoid contains a group of distinct protein kinase systems. A working model is accordingly proposed to interpret the interaction between these protein kinase systems.

Additional key words: dark-regulated protein phosphorylation; functional mass; light-dependent protein phosphorylation; light-harvesting complex 2; photosystem 2

Received: March 22, 2005; Accepted: July 15, 2005; Published: March 1, 2006  Show citation

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Lee, S.C., Chien, L.F., Van, R.C., Hsiao, Y.Y., Hong, J.L., & Pan, R.L. (2006). Radiation inactivation analysis of thylakoid protein kinase systems in light and in darkness. Photosynthetica, 44(1), 116-124. doi: 10.1007/s11099-005-0166-z
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