TY - JOUR
T1 - Chromatographic purification and determination of the carboxy-terminal sequences of photosystem II reaction center proteins, Dl and D2
AU - Takahashi, Yuichiro
AU - Nakane, Hiroyuki
AU - Kojima, Hisashi
AU - Satoh, Kimiyuki
N1 - Funding Information:
The authors thank Dr. S. Itoh of the National Institute for Basic Biology (NIBB) for amino acid analysis and valuable discussions. This work was supported in part by Grants-in-Aid for General Scientific Research and Scientific Research on Priority Area (nos. 63621003 and 01621003) from the Ministry of Education, Science and Culture of Japan, by the Itoh Science Foundation, by the CIBA-GEIGY Foundation, and by the NIBB Cooperative Research Program (88-104).
PY - 1990
Y1 - 1990
N2 - The Dl and D2 subunits of the reaction center of photosystem II are intrinsic proteins, each with a molecular mass of about 30 kDa. They exhibit considerable homology to each other in terms of primary structure. A procedure was developed for the separation and purification of these two proteins on a large scale from the photosystem II reaction center complex of spinach by high-performance liquid chromatography on a gel-permeation column in the presence of sodium dodecyl sulfate. The purification was achieved by a combination of two gel-permeation chromatographic steps performed with different concentrations of phosphate buffer, 200 mM and 50 mM, as the mobile phase. The purified Dl and D2 proteins were subjected to determination of their carboxy-terminal sequences by digestion of the proteins with carboxypeptidase Y. Comparison of the sequences deduced from the enzymatic analysis with the sequences deduced from the psb A and psb D genes of spinach indicates that the Dl protein ends at Ala-344 and the D2 protein at Leu-353. Thus, it appears that the Dl protein loses 9 amino acid residues from the carboxy-terminus, from Ala-345 to Gly-353, during maturation, while the D2 protein does not lose any amino acid residues from the carboxy-terminus.
AB - The Dl and D2 subunits of the reaction center of photosystem II are intrinsic proteins, each with a molecular mass of about 30 kDa. They exhibit considerable homology to each other in terms of primary structure. A procedure was developed for the separation and purification of these two proteins on a large scale from the photosystem II reaction center complex of spinach by high-performance liquid chromatography on a gel-permeation column in the presence of sodium dodecyl sulfate. The purification was achieved by a combination of two gel-permeation chromatographic steps performed with different concentrations of phosphate buffer, 200 mM and 50 mM, as the mobile phase. The purified Dl and D2 proteins were subjected to determination of their carboxy-terminal sequences by digestion of the proteins with carboxypeptidase Y. Comparison of the sequences deduced from the enzymatic analysis with the sequences deduced from the psb A and psb D genes of spinach indicates that the Dl protein ends at Ala-344 and the D2 protein at Leu-353. Thus, it appears that the Dl protein loses 9 amino acid residues from the carboxy-terminus, from Ala-345 to Gly-353, during maturation, while the D2 protein does not lose any amino acid residues from the carboxy-terminus.
KW - Carboxy-terminal sequence
KW - D2 protein
KW - Dl protein
KW - Gel permeation chromatography
KW - Photosynthesis
KW - Photosystem II
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M3 - Article
AN - SCOPUS:0001613533
VL - 31
SP - 273
EP - 280
JO - Plant and Cell Physiology
JF - Plant and Cell Physiology
SN - 0032-0781
IS - 2
ER -