Isolation and purification of a ubiquinone-cytochrome b-c1 complex from a photosynthetic bacterium, rhodopseudomonas sphaeroides

Ken Ichiro Takamiya, Michio Doi, Hideaki Okimatsu

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

A ubiquinone-cytochrome b-c1 complex was removed from chromatophore membranes of a Rhodopseudomonas sphaeroides green mutant by deoxycholate-cholate treatment of the chromatophores. The complex was purified by ammonium sulfate fractionation and gel filtration.The molecular weight of the purified complex was 240,000 (240 kD) and it was composed of seven subunits with molecular weights of 47 kD, 42 kD, 38 kD, 32 kD, 30 kD, 24 kD and 16 kD. The complex contained 1.54 and 3.42 nmol of cytochrome c1 and two different cytochrome b species per mg protein, respectively. It also contained 7.07 nmol of ubiquinone, 6.37 nmol of non-heme iron and about 3 nmol of carotenoids per mg protein. No flavins were detected. Heme staining indicated that the 32 kD-and 24 kD-subunits were cytochromes.The midpoint potential of cytochrome c1 was 245 mV, and the values for the cytochromes b were 60 mV and -75 mV at pH 7.2. The peak of the α-band of the reduced-minus-oxidized difference spectrum of cytochrome c1 was located at 552.5 nm, arid peaks of the b-type cytochromes with higher and lower midpoint potentials were located at 562 nm and 563 nm.The chemical and the subunit compositions of the purified complex reported here were similar to those obtained for the inner membranes of mitochondria of various organisms.

Original languageEnglish
Pages (from-to)987-997
Number of pages11
JournalPlant and Cell Physiology
Volume23
Issue number6
Publication statusPublished - Sep 1 1982

Fingerprint

Cytochromes c1
Photosynthetic Reaction Center Complex Proteins
Rhodobacter sphaeroides
photosynthetic bacteria
Cytochromes b
Ubiquinone
ubiquinones
cytochromes
cytochrome b
Chromatophores
Bacteria
chromatophores
Cytochrome b Group
Molecular Weight
Flavins
Cholates
Deoxycholic Acid
Membranes
deoxycholic acid
molecular weight

All Science Journal Classification (ASJC) codes

  • Physiology
  • Plant Science
  • Cell Biology

Cite this

Isolation and purification of a ubiquinone-cytochrome b-c1 complex from a photosynthetic bacterium, rhodopseudomonas sphaeroides. / Takamiya, Ken Ichiro; Doi, Michio; Okimatsu, Hideaki.

In: Plant and Cell Physiology, Vol. 23, No. 6, 01.09.1982, p. 987-997.

Research output: Contribution to journalArticle

@article{10cfcb3c9b0f4bd4b7426ad015fb6492,
title = "Isolation and purification of a ubiquinone-cytochrome b-c1 complex from a photosynthetic bacterium, rhodopseudomonas sphaeroides",
abstract = "A ubiquinone-cytochrome b-c1 complex was removed from chromatophore membranes of a Rhodopseudomonas sphaeroides green mutant by deoxycholate-cholate treatment of the chromatophores. The complex was purified by ammonium sulfate fractionation and gel filtration.The molecular weight of the purified complex was 240,000 (240 kD) and it was composed of seven subunits with molecular weights of 47 kD, 42 kD, 38 kD, 32 kD, 30 kD, 24 kD and 16 kD. The complex contained 1.54 and 3.42 nmol of cytochrome c1 and two different cytochrome b species per mg protein, respectively. It also contained 7.07 nmol of ubiquinone, 6.37 nmol of non-heme iron and about 3 nmol of carotenoids per mg protein. No flavins were detected. Heme staining indicated that the 32 kD-and 24 kD-subunits were cytochromes.The midpoint potential of cytochrome c1 was 245 mV, and the values for the cytochromes b were 60 mV and -75 mV at pH 7.2. The peak of the α-band of the reduced-minus-oxidized difference spectrum of cytochrome c1 was located at 552.5 nm, arid peaks of the b-type cytochromes with higher and lower midpoint potentials were located at 562 nm and 563 nm.The chemical and the subunit compositions of the purified complex reported here were similar to those obtained for the inner membranes of mitochondria of various organisms.",
author = "Takamiya, {Ken Ichiro} and Michio Doi and Hideaki Okimatsu",
year = "1982",
month = "9",
day = "1",
language = "English",
volume = "23",
pages = "987--997",
journal = "Plant and Cell Physiology",
issn = "0032-0781",
publisher = "Oxford University Press",
number = "6",

}

TY - JOUR

T1 - Isolation and purification of a ubiquinone-cytochrome b-c1 complex from a photosynthetic bacterium, rhodopseudomonas sphaeroides

AU - Takamiya, Ken Ichiro

AU - Doi, Michio

AU - Okimatsu, Hideaki

PY - 1982/9/1

Y1 - 1982/9/1

N2 - A ubiquinone-cytochrome b-c1 complex was removed from chromatophore membranes of a Rhodopseudomonas sphaeroides green mutant by deoxycholate-cholate treatment of the chromatophores. The complex was purified by ammonium sulfate fractionation and gel filtration.The molecular weight of the purified complex was 240,000 (240 kD) and it was composed of seven subunits with molecular weights of 47 kD, 42 kD, 38 kD, 32 kD, 30 kD, 24 kD and 16 kD. The complex contained 1.54 and 3.42 nmol of cytochrome c1 and two different cytochrome b species per mg protein, respectively. It also contained 7.07 nmol of ubiquinone, 6.37 nmol of non-heme iron and about 3 nmol of carotenoids per mg protein. No flavins were detected. Heme staining indicated that the 32 kD-and 24 kD-subunits were cytochromes.The midpoint potential of cytochrome c1 was 245 mV, and the values for the cytochromes b were 60 mV and -75 mV at pH 7.2. The peak of the α-band of the reduced-minus-oxidized difference spectrum of cytochrome c1 was located at 552.5 nm, arid peaks of the b-type cytochromes with higher and lower midpoint potentials were located at 562 nm and 563 nm.The chemical and the subunit compositions of the purified complex reported here were similar to those obtained for the inner membranes of mitochondria of various organisms.

AB - A ubiquinone-cytochrome b-c1 complex was removed from chromatophore membranes of a Rhodopseudomonas sphaeroides green mutant by deoxycholate-cholate treatment of the chromatophores. The complex was purified by ammonium sulfate fractionation and gel filtration.The molecular weight of the purified complex was 240,000 (240 kD) and it was composed of seven subunits with molecular weights of 47 kD, 42 kD, 38 kD, 32 kD, 30 kD, 24 kD and 16 kD. The complex contained 1.54 and 3.42 nmol of cytochrome c1 and two different cytochrome b species per mg protein, respectively. It also contained 7.07 nmol of ubiquinone, 6.37 nmol of non-heme iron and about 3 nmol of carotenoids per mg protein. No flavins were detected. Heme staining indicated that the 32 kD-and 24 kD-subunits were cytochromes.The midpoint potential of cytochrome c1 was 245 mV, and the values for the cytochromes b were 60 mV and -75 mV at pH 7.2. The peak of the α-band of the reduced-minus-oxidized difference spectrum of cytochrome c1 was located at 552.5 nm, arid peaks of the b-type cytochromes with higher and lower midpoint potentials were located at 562 nm and 563 nm.The chemical and the subunit compositions of the purified complex reported here were similar to those obtained for the inner membranes of mitochondria of various organisms.

UR - http://www.scopus.com/inward/record.url?scp=77957184771&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77957184771&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:77957184771

VL - 23

SP - 987

EP - 997

JO - Plant and Cell Physiology

JF - Plant and Cell Physiology

SN - 0032-0781

IS - 6

ER -