Site-directed mutagenesis of conserved histidines in the helix VIII domain of PsaB impairs assembly of the photosystem I reaction center without altering spectroscopic characteristics of P700

Liying Cui, Scott E. Bingham, Matthias Kuhn, Hanno Käss, Wolfgang Lubitz, Andrew Webber

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The chloroplast psaB gene encodes one of the polypeptides of the photosystem I reaction center heterodimer that coordinates the electron transfer components P700, A0, and A1. Histidine residues in the most highly conserved region of the PsaB protein are predicted to coordinate the P700 reaction center chlorophyll(s) and the initial electron acceptor, A0. Oligonucleotide-mediated site-directed mutagenesis and chloroplast transformation of Chlamydomonas reinhardtii have been used to determine the importance of these conserved histidines in photosystem I reaction center biogenesis and function. It is demonstrated that these histidine residues are essential for stable accumulation of the photosystem I reaction center. Protein pulse-labeling shows that changing the histidine residues impairs a post-translational step in reaction center assembly. Photosystem I complexes from the mutants have been characterized by Electron Nuclear Double Resonance and Electron Spin Echo Envelope Modulation spectroscopy to determine the impact of any mutations on P700 +. In all cases we determine that spectroscopic characteristics of P700 + remain unchanged. The implications of these results to current models of the photosystem I reaction center and related bacterial reaction centers are discussed.

Original languageEnglish (US)
Pages (from-to)1549-1558
Number of pages10
JournalBiochemistry
Volume34
Issue number5
StatePublished - 1995

Fingerprint

Photosystem I Protein Complex
Mutagenesis
Site-Directed Mutagenesis
Histidine
Electrons
Chloroplast Genes
Chlamydomonas reinhardtii
Electron Spin Resonance Spectroscopy
Chloroplasts
Chlorophyll
Oligonucleotides
Labeling
Spectrum Analysis
Proteins
Genes
Modulation
Spectroscopy
Peptides
Mutation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Site-directed mutagenesis of conserved histidines in the helix VIII domain of PsaB impairs assembly of the photosystem I reaction center without altering spectroscopic characteristics of P700 . / Cui, Liying; Bingham, Scott E.; Kuhn, Matthias; Käss, Hanno; Lubitz, Wolfgang; Webber, Andrew.

In: Biochemistry, Vol. 34, No. 5, 1995, p. 1549-1558.

Research output: Contribution to journalArticle

@article{70ce333eeb834bc2beb61bec1b1b6cf7,
title = "Site-directed mutagenesis of conserved histidines in the helix VIII domain of PsaB impairs assembly of the photosystem I reaction center without altering spectroscopic characteristics of P700",
abstract = "The chloroplast psaB gene encodes one of the polypeptides of the photosystem I reaction center heterodimer that coordinates the electron transfer components P700, A0, and A1. Histidine residues in the most highly conserved region of the PsaB protein are predicted to coordinate the P700 reaction center chlorophyll(s) and the initial electron acceptor, A0. Oligonucleotide-mediated site-directed mutagenesis and chloroplast transformation of Chlamydomonas reinhardtii have been used to determine the importance of these conserved histidines in photosystem I reaction center biogenesis and function. It is demonstrated that these histidine residues are essential for stable accumulation of the photosystem I reaction center. Protein pulse-labeling shows that changing the histidine residues impairs a post-translational step in reaction center assembly. Photosystem I complexes from the mutants have been characterized by Electron Nuclear Double Resonance and Electron Spin Echo Envelope Modulation spectroscopy to determine the impact of any mutations on P700 +. In all cases we determine that spectroscopic characteristics of P700 + remain unchanged. The implications of these results to current models of the photosystem I reaction center and related bacterial reaction centers are discussed.",
author = "Liying Cui and Bingham, {Scott E.} and Matthias Kuhn and Hanno K{\"a}ss and Wolfgang Lubitz and Andrew Webber",
year = "1995",
language = "English (US)",
volume = "34",
pages = "1549--1558",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "5",

}

TY - JOUR

T1 - Site-directed mutagenesis of conserved histidines in the helix VIII domain of PsaB impairs assembly of the photosystem I reaction center without altering spectroscopic characteristics of P700

AU - Cui, Liying

AU - Bingham, Scott E.

AU - Kuhn, Matthias

AU - Käss, Hanno

AU - Lubitz, Wolfgang

AU - Webber, Andrew

PY - 1995

Y1 - 1995

N2 - The chloroplast psaB gene encodes one of the polypeptides of the photosystem I reaction center heterodimer that coordinates the electron transfer components P700, A0, and A1. Histidine residues in the most highly conserved region of the PsaB protein are predicted to coordinate the P700 reaction center chlorophyll(s) and the initial electron acceptor, A0. Oligonucleotide-mediated site-directed mutagenesis and chloroplast transformation of Chlamydomonas reinhardtii have been used to determine the importance of these conserved histidines in photosystem I reaction center biogenesis and function. It is demonstrated that these histidine residues are essential for stable accumulation of the photosystem I reaction center. Protein pulse-labeling shows that changing the histidine residues impairs a post-translational step in reaction center assembly. Photosystem I complexes from the mutants have been characterized by Electron Nuclear Double Resonance and Electron Spin Echo Envelope Modulation spectroscopy to determine the impact of any mutations on P700 +. In all cases we determine that spectroscopic characteristics of P700 + remain unchanged. The implications of these results to current models of the photosystem I reaction center and related bacterial reaction centers are discussed.

AB - The chloroplast psaB gene encodes one of the polypeptides of the photosystem I reaction center heterodimer that coordinates the electron transfer components P700, A0, and A1. Histidine residues in the most highly conserved region of the PsaB protein are predicted to coordinate the P700 reaction center chlorophyll(s) and the initial electron acceptor, A0. Oligonucleotide-mediated site-directed mutagenesis and chloroplast transformation of Chlamydomonas reinhardtii have been used to determine the importance of these conserved histidines in photosystem I reaction center biogenesis and function. It is demonstrated that these histidine residues are essential for stable accumulation of the photosystem I reaction center. Protein pulse-labeling shows that changing the histidine residues impairs a post-translational step in reaction center assembly. Photosystem I complexes from the mutants have been characterized by Electron Nuclear Double Resonance and Electron Spin Echo Envelope Modulation spectroscopy to determine the impact of any mutations on P700 +. In all cases we determine that spectroscopic characteristics of P700 + remain unchanged. The implications of these results to current models of the photosystem I reaction center and related bacterial reaction centers are discussed.

UR - http://www.scopus.com/inward/record.url?scp=0028934118&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028934118&partnerID=8YFLogxK

M3 - Article

C2 - 7849014

AN - SCOPUS:0028934118

VL - 34

SP - 1549

EP - 1558

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 5

ER -