title: A genetically tagged Psb27 protein allows purification of two consecutive photosystem II (PSII) assembly intermediates in Synechocystis 6803, a cyanobacterium.
abstract: Photosystem II (PSII) is a large membrane bound molecular machine that catalyzes light-driven oxygen evolution from water. PSII constantly undergoes assembly and disassembly because of the unavoidable damage that results from its normal photochemistry. Thus, under physiological conditions, in addition to the active PSII complexes, there are always PSII subpopulations incompetent of oxygen evolution, but are in the process of undergoing elaborate biogenesis and repair. These transient complexes are difficult to characterize because of their low abundance, structural heterogeneity, and thermodynamic instability. In this study, we show that a genetically tagged Psb27 protein allows for the biochemical purification of two monomeric PSII assembly intermediates, one with an unprocessed form of D1 (His27ΔctpAPSII) and a second one with a mature form of D1 (His27PSII). Both forms were capable of light-induced charge separation, but unable to photooxidize water, largely because of the absence of a functional tetramanganese cluster. Unexpectedly, there was a significant amount of the extrinsic lumenal PsbO protein in the His27PSII, but not in the His27ΔctpAPSII complex. In contrast, two other lumenal proteins, PsbU and PsbV, were absent in both of these PSII intermediate complexes. Additionally, the only cytoplasmic extrinsic protein, Psb28 was detected in His27PSII complex. Based on these data, we have presented a refined model of PSII biogenesis, illustrating an important role of Psb27 as a gate-keeper during the complex assembly process of the oxygen-evolving centers in PSII.
authors: Department of Biology, Washington University, St. Louis, Missouri 63130, USA.
authors: Haijun
authors: H
authors: Liu
authors: Johnna L
authors: JL
authors: Roose
authors: Jeffrey C
authors: JC
authors: Cameron
authors: Himadri B
authors: HB
authors: Pakrasi
references: Proc Natl Acad Sci U S A. 2003 Jan 7;100(1):98-103
references: 12518057
references: Nat Struct Mol Biol. 2009 Mar;16(3):334-42
references: 19219048
references: Biochemistry. 2008 Nov 4;47(44):11637-46
references: 18834146
references: Biochemistry. 2002 Jun 25;41(25):8004-12
references: 12069591
references: Biochemistry. 2007 May 8;46(18):5542-51
references: 17432833
references: Plant Cell Physiol. 2002 Nov;43(11):1366-73
references: 12461137
references: J Photochem Photobiol B. 2011 Jul-Aug;104(1-2):204-11
references: 21382728
references: Nature. 2001 Feb 8;409(6821):739-43
references: 11217865
references: Biochim Biophys Acta. 1971 Nov 2;253(1):167-81
references: 5001676
references: Biotechniques. 1993 Nov;15(5):831-4
references: 8267974
references: Proc Natl Acad Sci U S A. 2001 Nov 6;98(23):13443-8
references: 11687660
references: Biochemistry. 1992 Jan 28;31(3):942-8
references: 1731951
references: Arch Biochem Biophys. 1987 Jul;256(1):295-301
references: 2440384
references: Plant Mol Biol. 2006 Jul;61(4-5):567-75
references: 16897475
references: Ann Bot. 2010 Jul;106(1):1-16
references: 20338950
references: Biochemistry. 1992 Jun 23;31(24):5616-20
references: 1610808
references: Nature. 2005 Dec 15;438(7070):1040-4
references: 16355230
references: J Photochem Photobiol B. 2011 Jul-Aug;104(1-2):191-203
references: 21349737
references: J Biol Chem. 2010 Jul 9;285(28):21391-8
references: 20444695
references: Science. 2004 Mar 19;303(5665):1831-8
references: 14764885
references: Plant Physiol. 2007 Dec;145(4):1361-70
references: 17921339
references: Biochemistry. 2008 Sep 16;47(37):9747-55
references: 18717592
references: Biochemistry. 2004 Mar 23;43(11):3152-66
references: 15023066
references: Anal Biochem. 1991 Dec;199(2):223-31
references: 1812789
references: Proc Natl Acad Sci U S A. 2007 Feb 13;104(7):2548-53
references: 17287351
references: Photosynth Res. 2001;70(3):245-7
references: 16252169
references: Eur J Cell Biol. 2010 Dec;89(12):974-82
references: 20943284
references: Plant Cell. 2006 Nov;18(11):3121-31
references: 17114356
references: Photosynth Res. 1990 Apr;24(1):1-13
references: 24419760
references: Electrophoresis. 2001 Apr;22(6):1004-7
references: 11358120
references: Biochemistry. 2009 Sep 22;48(37):8768-70
references: 19697957
references: J Biol Chem. 2008 Feb 15;283(7):4044-50
references: 18089572
references: Plant Physiol. 2009 Feb;149(2):1076-86
references: 19036835
references: Biochim Biophys Acta. 1998 Nov 2;1409(1):50-7
references: 9804889
references: J Am Chem Soc. 2010 Jan 20;132(2):446-7
references: 20038096
references: Biochem J. 2005 May 15;388(Pt 1):159-68
references: 15638811
references: J Biol Chem. 2004 Oct 29;279(44):45417-22
references: 15308630
references: J Phycol. 1968 Mar;4(1):1-4
references: 27067764