Determination of the PS I content of PS II core preparations using selective emission

A new emission of PS II at 780 nm

Jennifer Morton, Jeremy Hall, Paul Smith, Fusamichi Akita, Faisal Koua, Jian-Ren Shen, Elmars Krausz

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Routinely prepared PS II core samples are often contaminated by a significant (∼ 1-5%) fraction of PS I, as well as related proteins. This contamination is of little importance in many experiments, but masks the optical behaviour of the deep red state in PS II, which absorbs in the same spectral range (700-730 nm) as PS I (Hughes et al. 2006). When contamination levels are less than ~ 1%, it becomes difficult to quantify the PS I related components by gel-based, chromatographic, circular dichroism or EPR techniques. We have developed a fluorescence-based technique, taking advantage of the distinctively different low-temperature emission characteristics of PS II and PS I when excited near 700 nm. The approach has the advantage of providing the relative concentration of the two photosystems in a single spectral measurement. A sensitivity limit of 0.01% PS I (or better) can be achieved. The procedure is applied to PS II core preparations from spinach and Thermosynechococcus vulcanus. Measurements made of T. vulcanus PS II preparations prepared by re-dissolving crystallised material indicate a low but measurable PS I related content. The analysis provides strong evidence for a previously unreported fluorescence of PS II cores peaking near 780 nm. The excitation dependence of this emission as well as its appearance in both low PS I cyanobacterial and plant based PS II core preparations suggests its association with the deep red state of PS II.

Original languageEnglish
Pages (from-to)167-177
Number of pages11
JournalBiochimica et Biophysica Acta - Bioenergetics
Volume1837
Issue number1
DOIs
Publication statusPublished - 2014

Fingerprint

Contamination
Fluorescence
Core samples
Spinacia oleracea
Masks
Circular Dichroism
Paramagnetic resonance
Gels
Temperature
Proteins
Experiments
PS 5

Keywords

  • Fluorescence
  • Fluorescence line-narrowing
  • Photosystem I
  • Photosystem II
  • Spectral hole-burning

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology

Cite this

Determination of the PS I content of PS II core preparations using selective emission : A new emission of PS II at 780 nm. / Morton, Jennifer; Hall, Jeremy; Smith, Paul; Akita, Fusamichi; Koua, Faisal; Shen, Jian-Ren; Krausz, Elmars.

In: Biochimica et Biophysica Acta - Bioenergetics, Vol. 1837, No. 1, 2014, p. 167-177.

Research output: Contribution to journalArticle

@article{b169794853ec47489ae56482660bdd26,
title = "Determination of the PS I content of PS II core preparations using selective emission: A new emission of PS II at 780 nm",
abstract = "Routinely prepared PS II core samples are often contaminated by a significant (∼ 1-5{\%}) fraction of PS I, as well as related proteins. This contamination is of little importance in many experiments, but masks the optical behaviour of the deep red state in PS II, which absorbs in the same spectral range (700-730 nm) as PS I (Hughes et al. 2006). When contamination levels are less than ~ 1{\%}, it becomes difficult to quantify the PS I related components by gel-based, chromatographic, circular dichroism or EPR techniques. We have developed a fluorescence-based technique, taking advantage of the distinctively different low-temperature emission characteristics of PS II and PS I when excited near 700 nm. The approach has the advantage of providing the relative concentration of the two photosystems in a single spectral measurement. A sensitivity limit of 0.01{\%} PS I (or better) can be achieved. The procedure is applied to PS II core preparations from spinach and Thermosynechococcus vulcanus. Measurements made of T. vulcanus PS II preparations prepared by re-dissolving crystallised material indicate a low but measurable PS I related content. The analysis provides strong evidence for a previously unreported fluorescence of PS II cores peaking near 780 nm. The excitation dependence of this emission as well as its appearance in both low PS I cyanobacterial and plant based PS II core preparations suggests its association with the deep red state of PS II.",
keywords = "Fluorescence, Fluorescence line-narrowing, Photosystem I, Photosystem II, Spectral hole-burning",
author = "Jennifer Morton and Jeremy Hall and Paul Smith and Fusamichi Akita and Faisal Koua and Jian-Ren Shen and Elmars Krausz",
year = "2014",
doi = "10.1016/j.bbabio.2013.09.008",
language = "English",
volume = "1837",
pages = "167--177",
journal = "Biochimica et Biophysica Acta - Bioenergetics",
issn = "0005-2728",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Determination of the PS I content of PS II core preparations using selective emission

T2 - A new emission of PS II at 780 nm

AU - Morton, Jennifer

AU - Hall, Jeremy

AU - Smith, Paul

AU - Akita, Fusamichi

AU - Koua, Faisal

AU - Shen, Jian-Ren

AU - Krausz, Elmars

PY - 2014

Y1 - 2014

N2 - Routinely prepared PS II core samples are often contaminated by a significant (∼ 1-5%) fraction of PS I, as well as related proteins. This contamination is of little importance in many experiments, but masks the optical behaviour of the deep red state in PS II, which absorbs in the same spectral range (700-730 nm) as PS I (Hughes et al. 2006). When contamination levels are less than ~ 1%, it becomes difficult to quantify the PS I related components by gel-based, chromatographic, circular dichroism or EPR techniques. We have developed a fluorescence-based technique, taking advantage of the distinctively different low-temperature emission characteristics of PS II and PS I when excited near 700 nm. The approach has the advantage of providing the relative concentration of the two photosystems in a single spectral measurement. A sensitivity limit of 0.01% PS I (or better) can be achieved. The procedure is applied to PS II core preparations from spinach and Thermosynechococcus vulcanus. Measurements made of T. vulcanus PS II preparations prepared by re-dissolving crystallised material indicate a low but measurable PS I related content. The analysis provides strong evidence for a previously unreported fluorescence of PS II cores peaking near 780 nm. The excitation dependence of this emission as well as its appearance in both low PS I cyanobacterial and plant based PS II core preparations suggests its association with the deep red state of PS II.

AB - Routinely prepared PS II core samples are often contaminated by a significant (∼ 1-5%) fraction of PS I, as well as related proteins. This contamination is of little importance in many experiments, but masks the optical behaviour of the deep red state in PS II, which absorbs in the same spectral range (700-730 nm) as PS I (Hughes et al. 2006). When contamination levels are less than ~ 1%, it becomes difficult to quantify the PS I related components by gel-based, chromatographic, circular dichroism or EPR techniques. We have developed a fluorescence-based technique, taking advantage of the distinctively different low-temperature emission characteristics of PS II and PS I when excited near 700 nm. The approach has the advantage of providing the relative concentration of the two photosystems in a single spectral measurement. A sensitivity limit of 0.01% PS I (or better) can be achieved. The procedure is applied to PS II core preparations from spinach and Thermosynechococcus vulcanus. Measurements made of T. vulcanus PS II preparations prepared by re-dissolving crystallised material indicate a low but measurable PS I related content. The analysis provides strong evidence for a previously unreported fluorescence of PS II cores peaking near 780 nm. The excitation dependence of this emission as well as its appearance in both low PS I cyanobacterial and plant based PS II core preparations suggests its association with the deep red state of PS II.

KW - Fluorescence

KW - Fluorescence line-narrowing

KW - Photosystem I

KW - Photosystem II

KW - Spectral hole-burning

UR - http://www.scopus.com/inward/record.url?scp=84885351748&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84885351748&partnerID=8YFLogxK

U2 - 10.1016/j.bbabio.2013.09.008

DO - 10.1016/j.bbabio.2013.09.008

M3 - Article

VL - 1837

SP - 167

EP - 177

JO - Biochimica et Biophysica Acta - Bioenergetics

JF - Biochimica et Biophysica Acta - Bioenergetics

SN - 0005-2728

IS - 1

ER -