Details
| Original language | English |
|---|---|
| Pages (from-to) | 1683-1692 |
| Number of pages | 10 |
| Journal | PHYTOCHEMISTRY |
| Volume | 65 |
| Issue number | 12 |
| Publication status | Published - 21 May 2004 |
Abstract
A project to investigate the supramolecular structure of photosystems was initiated, which is based on protein solubilizations by digitonin, protein separations by Blue native (BN)-polyacrylamide gel electrophoresis (PAGE) and protein identifications by mass spectrometry (MS). Under the conditions applied, nine photosystem supercomplexes could be described for chloroplasts of Arabidopsis, which have apparent molecular masses between 600 and 3200 kDa on BN gels. Identities of the supercomplexes were determined on the basis of their subunit compositions as documented by 2D BN/SDS-PAGE and BN/BN-PAGE. Two supercomplexes of 1060 and ∼1600 kDa represent dimeric and trimeric forms of photosystem I (PSI), which include tightly bound LHCI proteins. Compared to monomeric PSI, these protein complexes are of low abundance. In contrast, photosystem II mainly forms part of dominant supercomplexes of 850, 1000, 1050 and 1300 kDa. According to our interpretation, these supercomplexes contain dimeric PSII, 1-4 LHCII trimers and additionally monomeric LHCII proteins. The 1300-kDa PSII supercomplex (containing four LHCII trimers) is partially converted into the 1000-kDa PSII supercomplex (containing two LHCII trimers) in the presence of dodecylmaltoside on 2D BN/BN gels. Analyses of peptides of the trypsinated 1300-kDa PSII supercomplex by mass spectrometry allowed to identify known subunits of the PSII core complex and additionally LHCII proteins encoded by eight different genes in Arabidopsis. Further application of this experimental approach will allow new insights into the supermolecular organization of photosystems in plants.
Keywords
- Arabidopsis thaliana, bf complex, Blue-native polyacrylamide gel electrophoresis, Chloroplasts, Light harvesting complex, Mass spectrometry, Photosystem I, Photosystem II, Proteomics, Supercomplexes
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Agricultural and Biological Sciences(all)
- Plant Science
- Agricultural and Biological Sciences(all)
- Horticulture
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In: PHYTOCHEMISTRY, Vol. 65, No. 12, 21.05.2004, p. 1683-1692.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Proteomic approach to characterize the supramolecular organization of photosystems in higher plants
AU - Heinemeyer, Jesco
AU - Eubel, Holger
AU - Wehmhöner, Dirk
AU - Jänsch, Lothar
AU - Braun, Hans Peter
N1 - Funding information: We wish to thank Dagmar Lewejohann for the cultivation of Arabidopsis suspension cell cultures and expert technical assistance. The present work was supported by the Deutsche Forschungsgemeinschaft.
PY - 2004/5/21
Y1 - 2004/5/21
N2 - A project to investigate the supramolecular structure of photosystems was initiated, which is based on protein solubilizations by digitonin, protein separations by Blue native (BN)-polyacrylamide gel electrophoresis (PAGE) and protein identifications by mass spectrometry (MS). Under the conditions applied, nine photosystem supercomplexes could be described for chloroplasts of Arabidopsis, which have apparent molecular masses between 600 and 3200 kDa on BN gels. Identities of the supercomplexes were determined on the basis of their subunit compositions as documented by 2D BN/SDS-PAGE and BN/BN-PAGE. Two supercomplexes of 1060 and ∼1600 kDa represent dimeric and trimeric forms of photosystem I (PSI), which include tightly bound LHCI proteins. Compared to monomeric PSI, these protein complexes are of low abundance. In contrast, photosystem II mainly forms part of dominant supercomplexes of 850, 1000, 1050 and 1300 kDa. According to our interpretation, these supercomplexes contain dimeric PSII, 1-4 LHCII trimers and additionally monomeric LHCII proteins. The 1300-kDa PSII supercomplex (containing four LHCII trimers) is partially converted into the 1000-kDa PSII supercomplex (containing two LHCII trimers) in the presence of dodecylmaltoside on 2D BN/BN gels. Analyses of peptides of the trypsinated 1300-kDa PSII supercomplex by mass spectrometry allowed to identify known subunits of the PSII core complex and additionally LHCII proteins encoded by eight different genes in Arabidopsis. Further application of this experimental approach will allow new insights into the supermolecular organization of photosystems in plants.
AB - A project to investigate the supramolecular structure of photosystems was initiated, which is based on protein solubilizations by digitonin, protein separations by Blue native (BN)-polyacrylamide gel electrophoresis (PAGE) and protein identifications by mass spectrometry (MS). Under the conditions applied, nine photosystem supercomplexes could be described for chloroplasts of Arabidopsis, which have apparent molecular masses between 600 and 3200 kDa on BN gels. Identities of the supercomplexes were determined on the basis of their subunit compositions as documented by 2D BN/SDS-PAGE and BN/BN-PAGE. Two supercomplexes of 1060 and ∼1600 kDa represent dimeric and trimeric forms of photosystem I (PSI), which include tightly bound LHCI proteins. Compared to monomeric PSI, these protein complexes are of low abundance. In contrast, photosystem II mainly forms part of dominant supercomplexes of 850, 1000, 1050 and 1300 kDa. According to our interpretation, these supercomplexes contain dimeric PSII, 1-4 LHCII trimers and additionally monomeric LHCII proteins. The 1300-kDa PSII supercomplex (containing four LHCII trimers) is partially converted into the 1000-kDa PSII supercomplex (containing two LHCII trimers) in the presence of dodecylmaltoside on 2D BN/BN gels. Analyses of peptides of the trypsinated 1300-kDa PSII supercomplex by mass spectrometry allowed to identify known subunits of the PSII core complex and additionally LHCII proteins encoded by eight different genes in Arabidopsis. Further application of this experimental approach will allow new insights into the supermolecular organization of photosystems in plants.
KW - Arabidopsis thaliana
KW - bf complex
KW - Blue-native polyacrylamide gel electrophoresis
KW - Chloroplasts
KW - Light harvesting complex
KW - Mass spectrometry
KW - Photosystem I
KW - Photosystem II
KW - Proteomics
KW - Supercomplexes
UR - http://www.scopus.com/inward/record.url?scp=3242754429&partnerID=8YFLogxK
U2 - 10.1016/j.phytochem.2004.04.022
DO - 10.1016/j.phytochem.2004.04.022
M3 - Article
C2 - 15276430
AN - SCOPUS:3242754429
VL - 65
SP - 1683
EP - 1692
JO - PHYTOCHEMISTRY
JF - PHYTOCHEMISTRY
SN - 0031-9422
IS - 12
ER -