Proteomic analysis of a highly active photosystem II preparation from the cyanobacterium Synechocystis sp. PCC 6803 reveals the presence of novel polypeptides.
A highly active oxygen-evolving photosystem II (PSII) complex was purified from the HT-3 strain of the widely used cyanobacterium Synechocystis sp. PCC 6803, in which the CP47 polypeptide has been genetically engineered to contain a polyhistidine tag at its carboxyl terminus [Bricker, T. M., Morvant, J., Masri, N., Sutton, H. M., and Frankel, L. K. (1998) Biochim. Biophys. Acta 1409, 50-57]. These purified PSII centers had four manganese atoms, one calcium atom, and two cytochrome b(559) hemes each. Optical absorption and fluorescence emission spectroscopy as well as western immunoblot analysis demonstrated that the purified PSII preparation was devoid of any contamination with photosystem I and phycobiliproteins. A comprehensive proteomic analysis using a system designed to enhance resolution of low-molecular-weight polypeptides, followed by MALDI mass spectrometry and N-terminal amino acid sequencing, identified 31 distinct polypeptides in this PSII preparation. We propose a new nomenclature for the polypeptide components of PSII identified after PsbZ, which proceeds sequentially from Psb27. During this study, the polypeptides PsbJ, PsbM, PsbX, PsbY, PsbZ, Psb27, and Psb28 proteins were detected for the first time in a purified PSII complex from Synechocystis 6803. Five novel polypeptides were also identified in this preparation. They included the Sll1638 protein, which shares significant sequence similarity to PsbQ, a peripheral protein of PSII that was previously thought to be present only in chloroplasts. This work describes newly identified proteins in a highly purified cyanobacterial PSII preparation that is being widely used to investigate the structure, function, and biogenesis of this photosystem.
Kashino
Yasuhiro
Y
Department of Biology, Washington University, One Brookings Drive, St. Louis, MO 63130, USA.
Lauber
Wendy M
WM
Carroll
James A
JA
Wang
Qingjun
Q
Whitmarsh
John
J
Satoh
Kazuhiko
K
Pakrasi
Himadri B
HB
D029681:Arabidopsis Proteins; D001426:Bacterial Proteins; D016227:Benzoquinones / Q000378:metabolism; D002118:Calcium / Q000032:analysis; D002734:Chlorophyll / Q000032:analysis; D000458:Cyanobacteria / Q000737:chemistry* / Q000201:enzymology; D003573:Cytochrome b Group / Q000032:analysis; D003574:Cytochrome c Group / Q000302:isolation & purification; D007501:Iron / Q000032:analysis; D045342:Light-Harvesting Protein Complexes; D008345:Manganese / Q000032:analysis; D008970:Molecular Weight; D010100:Oxygen / Q000378:metabolism; D010446:Peptide Fragments / Q000302:isolation & purification*; D010674:Pheophytins / Q000032:analysis; D045322:Photosynthetic Reaction Center Complex Proteins / Q000032:analysis* / Q000302:isolation & purification; D045331:Photosystem I Protein Complex; D045332:Photosystem II Protein Complex; D045524:Phycobilisomes; D010971:Plastoquinone / Q000032:analysis; D011506:Proteins / Q000302:isolation & purification; D020543:Proteome / Q000032:analysis*; D013050:Spectrometry, Fluorescence; D020524:Thylakoids / Q000737:chemistry; D024341:Xanthophylls; D065146:Zeaxanthins; D019207:beta Carotene / Q000031:analogs & derivatives / Q000032:analysis
D029681:Arabidopsis Proteins; D001426:Bacterial Proteins; D016227:Benzoquinones; D003573:Cytochrome b Group; D003574:Cytochrome c Group; D045342:Light-Harvesting Protein Complexes; D010446:Peptide Fragments; D010674:Pheophytins; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D045332:Photosystem II Protein Complex; D045524:Phycobilisomes; D011506:Proteins; D020543:Proteome; C107126:PsbU protein, Synechocystis; D024341:Xanthophylls; D065146:Zeaxanthins; C054266:photosystem II manganese-stabilizing protein; C121497:psbQ protein, Arabidopsis; D019207:beta Carotene; D002734:Chlorophyll; D008345:Manganese; C071926:2,6-dichlorobenzoquinone; C019313:cytochrome b559; C031732:cytochrome C-550; D007501:Iron; D010971:Plastoquinone; D010100:Oxygen; D002118:Calcium