Characterization of a resolved oxygen-evolving Photosystem II preparation from spinach thylakoids

Eric Lam, Barbara Baltimore, William Ortiz, Susan Chollar, Anastasios Melis, Richard Malkin

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

An oxygen-evolving Photosystem (PS) II preparation was isolated after Triton X-100 treatment of spinach thylakoids in the presence of Mg2+. The structural and functional components of this preparation have been identified by SDS-polyacrylamide gel electrophoresis and sensitive spectrophotometric analysis. The main findings were: (1) The concentration of the primary acceptor Q of PS II was 1 per 230 chlorophyll molecules. (2) There are 6 to 7 plastoquinone molecules associated with a 'quinone-pool' reducible by Q. (3) The only cytochrome present in significant amounts (cytochrome b-559) occurred at a concentration of 1 per 125 chlorophyll molecules. (4) The only kind of photochemical reaction center complex present was identified by fluorescence induction kinetic analysis as PS IIα. (5) An Em = - 10 mV has been measured at pH 7.8 for the primary electron acceptor Qα of PS IIα. (6) With conventional SDS-polyacrylamide gel electrophoresis, the preparation was resolved into 13 prominent polypeptide bands with relative molecular masses of 63, 55, 51, 48, 37, 33, 28, 27, 25, 22, 15, 13 and 10 kDa. The 28 kDa band was identified as the PS II light-harvesting chlorophyll a b-protein. In the presence of 2 M urea, however, SDS-polyacrylamide gel electrophoresis showed seven prominent polypeptides with molecular masses of 47, 39, 31, 29, 27, 26 and 13 kDa as well as several minor components. CP I under identical conditions had a molecular mass of 60-63 kDa.

Original languageEnglish (US)
Pages (from-to)201-211
Number of pages11
JournalBBA - Bioenergetics
Volume724
Issue number2
DOIs
StatePublished - Aug 31 1983
Externally publishedYes

Fingerprint

Thylakoids
Photosystem II Protein Complex
Spinacia oleracea
Oxygen
Molecular mass
Electrophoresis
Polyacrylamide Gel Electrophoresis
Chlorophyll
Molecules
Plastoquinone
Chlorophyll Binding Proteins
Peptides
Photochemical reactions
Octoxynol
Cytochromes
Urea
Fluorescence
Electrons
Light
Kinetics

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Cell Biology

Cite this

Lam, Eric ; Baltimore, Barbara ; Ortiz, William ; Chollar, Susan ; Melis, Anastasios ; Malkin, Richard. / Characterization of a resolved oxygen-evolving Photosystem II preparation from spinach thylakoids. In: BBA - Bioenergetics. 1983 ; Vol. 724, No. 2. pp. 201-211.
@article{518556495f7049e49662ea90daea38bf,
title = "Characterization of a resolved oxygen-evolving Photosystem II preparation from spinach thylakoids",
abstract = "An oxygen-evolving Photosystem (PS) II preparation was isolated after Triton X-100 treatment of spinach thylakoids in the presence of Mg2+. The structural and functional components of this preparation have been identified by SDS-polyacrylamide gel electrophoresis and sensitive spectrophotometric analysis. The main findings were: (1) The concentration of the primary acceptor Q of PS II was 1 per 230 chlorophyll molecules. (2) There are 6 to 7 plastoquinone molecules associated with a 'quinone-pool' reducible by Q. (3) The only cytochrome present in significant amounts (cytochrome b-559) occurred at a concentration of 1 per 125 chlorophyll molecules. (4) The only kind of photochemical reaction center complex present was identified by fluorescence induction kinetic analysis as PS IIα. (5) An Em = - 10 mV has been measured at pH 7.8 for the primary electron acceptor Qα of PS IIα. (6) With conventional SDS-polyacrylamide gel electrophoresis, the preparation was resolved into 13 prominent polypeptide bands with relative molecular masses of 63, 55, 51, 48, 37, 33, 28, 27, 25, 22, 15, 13 and 10 kDa. The 28 kDa band was identified as the PS II light-harvesting chlorophyll a b-protein. In the presence of 2 M urea, however, SDS-polyacrylamide gel electrophoresis showed seven prominent polypeptides with molecular masses of 47, 39, 31, 29, 27, 26 and 13 kDa as well as several minor components. CP I under identical conditions had a molecular mass of 60-63 kDa.",
author = "Eric Lam and Barbara Baltimore and William Ortiz and Susan Chollar and Anastasios Melis and Richard Malkin",
year = "1983",
month = "8",
day = "31",
doi = "https://doi.org/10.1016/0005-2728(83)90139-1",
language = "English (US)",
volume = "724",
pages = "201--211",
journal = "BBA - Bioenergetics",
issn = "0005-2728",
publisher = "Elsevier",
number = "2",

}

Characterization of a resolved oxygen-evolving Photosystem II preparation from spinach thylakoids. / Lam, Eric; Baltimore, Barbara; Ortiz, William; Chollar, Susan; Melis, Anastasios; Malkin, Richard.

In: BBA - Bioenergetics, Vol. 724, No. 2, 31.08.1983, p. 201-211.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Characterization of a resolved oxygen-evolving Photosystem II preparation from spinach thylakoids

AU - Lam, Eric

AU - Baltimore, Barbara

AU - Ortiz, William

AU - Chollar, Susan

AU - Melis, Anastasios

AU - Malkin, Richard

PY - 1983/8/31

Y1 - 1983/8/31

N2 - An oxygen-evolving Photosystem (PS) II preparation was isolated after Triton X-100 treatment of spinach thylakoids in the presence of Mg2+. The structural and functional components of this preparation have been identified by SDS-polyacrylamide gel electrophoresis and sensitive spectrophotometric analysis. The main findings were: (1) The concentration of the primary acceptor Q of PS II was 1 per 230 chlorophyll molecules. (2) There are 6 to 7 plastoquinone molecules associated with a 'quinone-pool' reducible by Q. (3) The only cytochrome present in significant amounts (cytochrome b-559) occurred at a concentration of 1 per 125 chlorophyll molecules. (4) The only kind of photochemical reaction center complex present was identified by fluorescence induction kinetic analysis as PS IIα. (5) An Em = - 10 mV has been measured at pH 7.8 for the primary electron acceptor Qα of PS IIα. (6) With conventional SDS-polyacrylamide gel electrophoresis, the preparation was resolved into 13 prominent polypeptide bands with relative molecular masses of 63, 55, 51, 48, 37, 33, 28, 27, 25, 22, 15, 13 and 10 kDa. The 28 kDa band was identified as the PS II light-harvesting chlorophyll a b-protein. In the presence of 2 M urea, however, SDS-polyacrylamide gel electrophoresis showed seven prominent polypeptides with molecular masses of 47, 39, 31, 29, 27, 26 and 13 kDa as well as several minor components. CP I under identical conditions had a molecular mass of 60-63 kDa.

AB - An oxygen-evolving Photosystem (PS) II preparation was isolated after Triton X-100 treatment of spinach thylakoids in the presence of Mg2+. The structural and functional components of this preparation have been identified by SDS-polyacrylamide gel electrophoresis and sensitive spectrophotometric analysis. The main findings were: (1) The concentration of the primary acceptor Q of PS II was 1 per 230 chlorophyll molecules. (2) There are 6 to 7 plastoquinone molecules associated with a 'quinone-pool' reducible by Q. (3) The only cytochrome present in significant amounts (cytochrome b-559) occurred at a concentration of 1 per 125 chlorophyll molecules. (4) The only kind of photochemical reaction center complex present was identified by fluorescence induction kinetic analysis as PS IIα. (5) An Em = - 10 mV has been measured at pH 7.8 for the primary electron acceptor Qα of PS IIα. (6) With conventional SDS-polyacrylamide gel electrophoresis, the preparation was resolved into 13 prominent polypeptide bands with relative molecular masses of 63, 55, 51, 48, 37, 33, 28, 27, 25, 22, 15, 13 and 10 kDa. The 28 kDa band was identified as the PS II light-harvesting chlorophyll a b-protein. In the presence of 2 M urea, however, SDS-polyacrylamide gel electrophoresis showed seven prominent polypeptides with molecular masses of 47, 39, 31, 29, 27, 26 and 13 kDa as well as several minor components. CP I under identical conditions had a molecular mass of 60-63 kDa.

UR - http://www.scopus.com/inward/record.url?scp=0000078233&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0000078233&partnerID=8YFLogxK

U2 - https://doi.org/10.1016/0005-2728(83)90139-1

DO - https://doi.org/10.1016/0005-2728(83)90139-1

M3 - Article

VL - 724

SP - 201

EP - 211

JO - BBA - Bioenergetics

JF - BBA - Bioenergetics

SN - 0005-2728

IS - 2

ER -