Transmembrane orientation of reaction centers in proteoliposomes from Rhodopseudomonas sphaeroides

Koh Iba, Ken Ichiro Takamiya, Hiroyuki Arata, Yoshihiro Toh, Mitsuo Nishimura

研究成果: ジャーナルへの寄稿記事

6 引用 (Scopus)

抄録

The photochemical reaction centers from Rhodopseudomonas sphaeroides were reconstituted with soybean phospholipids into liposomes by the cholate-dialysis method. The transmembrane orientation of the reaction centers in the proteoliposomes and the morphology of the vesicles were investigated. The orientastion was determined by the reduction of externally added cytochrome c after its photo-oxidation by a flash. The structure of the vesicles was examined by electron microscope.Discontinuous sucrose density gradient centrifugation yielded several proteoli-posome fractions with different vesicular sizes and reaction-center orientations. The proportion of the reaction centers that exposed their cytochrome c reacting sites to the outside of the vesicles increased from 45 to 85% with an increase of the vesicular size. The proportion also depended on the ionic composition of the dialysis buffer. The optimal ionic environment during the dialysis (100 mM NaCl or 2.5 mM MgSO4 gave a liposome yield of 25-30% with a highly asymmetric orientation (>60%). Entrapping of cytochrome c molecules into the phospholipid vesicles had little effect on the orientation of the reaction centers.

元の言語英語
ページ(範囲)1823-1830
ページ数8
ジャーナルJournal of biochemistry
96
発行部数6
DOI
出版物ステータス出版済み - 1 1 1984

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Rhodobacter sphaeroides
Dialysis
Cytochromes c
Liposomes
Phospholipids
Cholates
Photooxidation
Density Gradient Centrifugation
Centrifugation
Photochemical reactions
Soybeans
Sucrose
Buffers
Electron microscopes
Electrons
Molecules
Chemical analysis
proteoliposomes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

これを引用

Transmembrane orientation of reaction centers in proteoliposomes from Rhodopseudomonas sphaeroides. / Iba, Koh; Takamiya, Ken Ichiro; Arata, Hiroyuki; Toh, Yoshihiro; Nishimura, Mitsuo.

:: Journal of biochemistry, 巻 96, 番号 6, 01.01.1984, p. 1823-1830.

研究成果: ジャーナルへの寄稿記事

Iba, Koh ; Takamiya, Ken Ichiro ; Arata, Hiroyuki ; Toh, Yoshihiro ; Nishimura, Mitsuo. / Transmembrane orientation of reaction centers in proteoliposomes from Rhodopseudomonas sphaeroides. :: Journal of biochemistry. 1984 ; 巻 96, 番号 6. pp. 1823-1830.
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abstract = "The photochemical reaction centers from Rhodopseudomonas sphaeroides were reconstituted with soybean phospholipids into liposomes by the cholate-dialysis method. The transmembrane orientation of the reaction centers in the proteoliposomes and the morphology of the vesicles were investigated. The orientastion was determined by the reduction of externally added cytochrome c after its photo-oxidation by a flash. The structure of the vesicles was examined by electron microscope.Discontinuous sucrose density gradient centrifugation yielded several proteoli-posome fractions with different vesicular sizes and reaction-center orientations. The proportion of the reaction centers that exposed their cytochrome c reacting sites to the outside of the vesicles increased from 45 to 85{\%} with an increase of the vesicular size. The proportion also depended on the ionic composition of the dialysis buffer. The optimal ionic environment during the dialysis (100 mM NaCl or 2.5 mM MgSO4 gave a liposome yield of 25-30{\%} with a highly asymmetric orientation (>60{\%}). Entrapping of cytochrome c molecules into the phospholipid vesicles had little effect on the orientation of the reaction centers.",
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AU - Nishimura, Mitsuo

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AB - The photochemical reaction centers from Rhodopseudomonas sphaeroides were reconstituted with soybean phospholipids into liposomes by the cholate-dialysis method. The transmembrane orientation of the reaction centers in the proteoliposomes and the morphology of the vesicles were investigated. The orientastion was determined by the reduction of externally added cytochrome c after its photo-oxidation by a flash. The structure of the vesicles was examined by electron microscope.Discontinuous sucrose density gradient centrifugation yielded several proteoli-posome fractions with different vesicular sizes and reaction-center orientations. The proportion of the reaction centers that exposed their cytochrome c reacting sites to the outside of the vesicles increased from 45 to 85% with an increase of the vesicular size. The proportion also depended on the ionic composition of the dialysis buffer. The optimal ionic environment during the dialysis (100 mM NaCl or 2.5 mM MgSO4 gave a liposome yield of 25-30% with a highly asymmetric orientation (>60%). Entrapping of cytochrome c molecules into the phospholipid vesicles had little effect on the orientation of the reaction centers.

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