Spectroscopic evidence from site-directed mutants of Synechocystis PCC6803 in favor of a close interaction between histidine 189 and redox-active tyrosine 160, both of polypeptide D2 of the photosystem II reaction center

X. S. Tang, D. A. Chisholm, G Charles Dismukes, Gary W Brudvig, B. A. Diner

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Abstract

The reaction center of photosystem II of oxygenic photosynthesis contains two redox-active tyrosines called Z and D, each of which can act as an electron donor to the oxidized primary electron donor, P680 +. These tyrosines are located in homologous positions on the third transmembrane α- helix of each of the two homologous polypeptides, D1 and D2, that comprise the reaction center. Tyrosine D of polypeptide D2 has been proposed, upon oxidation, to give up its phenolic proton to a nearby basic amino acid residue, forming a neutral radical. Modeling studies have pointed to His190 (spinach numbering) as a likely candidate for this basic residue. As a test of this hypothesis, we have constructed three site-directed mutations in the D2 polypeptide of the cyanobacterium Synechocystis sp. PCC6803. His189 (the Synechocystis homologue of His190 of spinach) has been replaced by glutamine, aspartate, or leucine. Instead of the normal D· EPR signal (g = 2.0046; line width 16-19 G), PSII core complexes isolated from these three mutants show an altered dark-stable EPR signal with a narrowed line width (11-13 G), and g values of 2.0046, 2.0043, and 2.0042 for the His189Gln, His189Asp, and His189Leu mutants, respectively. Despite the reduced line width, these EPR signals show g values and microwave-power saturation properties similar to the normal D· signal. Furthermore, specific deuteration in one of those mutants at the 3 and 5 positions of the phenol ring of the photosystem II reaction center tyrosines results in a loss of hyperfine structure of the EPR signal, proving that the signal indeed arises from tyrosine. Proton-ENDOR studies of these tyrosine radicals show that one hyperfine coupling component of 3.5-3.6 MHz, observed in the wild-type strain disappears in all three mutants. Upon incubation of wild-type photosystem II core complexes in D2O, this hyperfine coupling is lost, indicating that it originates from an exchangeable proton, most likely interacting with D· through a hydrogen bond. These results provide strong experimental evidence in favor of a close interaction between His189 and Tyr160 in the D2 polypeptide of photosystem II. This observation provides support for a model in which an imidazole nitrogen of His189 accepts the phenolic proton of Tyr160 upon oxidation of D, forming a back hydrogen bond to the phenolic oxygen of the neutral tyrosyl radical.

Original languageEnglish
Pages (from-to)13742-13748
Number of pages7
JournalBiochemistry
Volume32
Issue number49
DOIs
Publication statusPublished - 1993

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Synechocystis
Photosystem II Protein Complex
Histidine
Oxidation-Reduction
Tyrosine
Paramagnetic resonance
Protons
Peptides
Linewidth
Spinacia oleracea
Hydrogen
Hydrogen bonds
Electrons
Basic Amino Acids
Oxidation
Photosynthesis
Electron Spin Resonance Spectroscopy
Cyanobacteria
Microwaves
Phenol

ASJC Scopus subject areas

  • Biochemistry

Cite this

@article{01968759cf094edd9115b8ed399d1e05,
title = "Spectroscopic evidence from site-directed mutants of Synechocystis PCC6803 in favor of a close interaction between histidine 189 and redox-active tyrosine 160, both of polypeptide D2 of the photosystem II reaction center",
abstract = "The reaction center of photosystem II of oxygenic photosynthesis contains two redox-active tyrosines called Z and D, each of which can act as an electron donor to the oxidized primary electron donor, P680 +. These tyrosines are located in homologous positions on the third transmembrane α- helix of each of the two homologous polypeptides, D1 and D2, that comprise the reaction center. Tyrosine D of polypeptide D2 has been proposed, upon oxidation, to give up its phenolic proton to a nearby basic amino acid residue, forming a neutral radical. Modeling studies have pointed to His190 (spinach numbering) as a likely candidate for this basic residue. As a test of this hypothesis, we have constructed three site-directed mutations in the D2 polypeptide of the cyanobacterium Synechocystis sp. PCC6803. His189 (the Synechocystis homologue of His190 of spinach) has been replaced by glutamine, aspartate, or leucine. Instead of the normal D· EPR signal (g = 2.0046; line width 16-19 G), PSII core complexes isolated from these three mutants show an altered dark-stable EPR signal with a narrowed line width (11-13 G), and g values of 2.0046, 2.0043, and 2.0042 for the His189Gln, His189Asp, and His189Leu mutants, respectively. Despite the reduced line width, these EPR signals show g values and microwave-power saturation properties similar to the normal D· signal. Furthermore, specific deuteration in one of those mutants at the 3 and 5 positions of the phenol ring of the photosystem II reaction center tyrosines results in a loss of hyperfine structure of the EPR signal, proving that the signal indeed arises from tyrosine. Proton-ENDOR studies of these tyrosine radicals show that one hyperfine coupling component of 3.5-3.6 MHz, observed in the wild-type strain disappears in all three mutants. Upon incubation of wild-type photosystem II core complexes in D2O, this hyperfine coupling is lost, indicating that it originates from an exchangeable proton, most likely interacting with D· through a hydrogen bond. These results provide strong experimental evidence in favor of a close interaction between His189 and Tyr160 in the D2 polypeptide of photosystem II. This observation provides support for a model in which an imidazole nitrogen of His189 accepts the phenolic proton of Tyr160 upon oxidation of D, forming a back hydrogen bond to the phenolic oxygen of the neutral tyrosyl radical.",
author = "Tang, {X. S.} and Chisholm, {D. A.} and Dismukes, {G Charles} and Brudvig, {Gary W} and Diner, {B. A.}",
year = "1993",
doi = "10.1021/bi00212a045",
language = "English",
volume = "32",
pages = "13742--13748",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "49",

}

TY - JOUR

T1 - Spectroscopic evidence from site-directed mutants of Synechocystis PCC6803 in favor of a close interaction between histidine 189 and redox-active tyrosine 160, both of polypeptide D2 of the photosystem II reaction center

AU - Tang, X. S.

AU - Chisholm, D. A.

AU - Dismukes, G Charles

AU - Brudvig, Gary W

AU - Diner, B. A.

PY - 1993

Y1 - 1993

N2 - The reaction center of photosystem II of oxygenic photosynthesis contains two redox-active tyrosines called Z and D, each of which can act as an electron donor to the oxidized primary electron donor, P680 +. These tyrosines are located in homologous positions on the third transmembrane α- helix of each of the two homologous polypeptides, D1 and D2, that comprise the reaction center. Tyrosine D of polypeptide D2 has been proposed, upon oxidation, to give up its phenolic proton to a nearby basic amino acid residue, forming a neutral radical. Modeling studies have pointed to His190 (spinach numbering) as a likely candidate for this basic residue. As a test of this hypothesis, we have constructed three site-directed mutations in the D2 polypeptide of the cyanobacterium Synechocystis sp. PCC6803. His189 (the Synechocystis homologue of His190 of spinach) has been replaced by glutamine, aspartate, or leucine. Instead of the normal D· EPR signal (g = 2.0046; line width 16-19 G), PSII core complexes isolated from these three mutants show an altered dark-stable EPR signal with a narrowed line width (11-13 G), and g values of 2.0046, 2.0043, and 2.0042 for the His189Gln, His189Asp, and His189Leu mutants, respectively. Despite the reduced line width, these EPR signals show g values and microwave-power saturation properties similar to the normal D· signal. Furthermore, specific deuteration in one of those mutants at the 3 and 5 positions of the phenol ring of the photosystem II reaction center tyrosines results in a loss of hyperfine structure of the EPR signal, proving that the signal indeed arises from tyrosine. Proton-ENDOR studies of these tyrosine radicals show that one hyperfine coupling component of 3.5-3.6 MHz, observed in the wild-type strain disappears in all three mutants. Upon incubation of wild-type photosystem II core complexes in D2O, this hyperfine coupling is lost, indicating that it originates from an exchangeable proton, most likely interacting with D· through a hydrogen bond. These results provide strong experimental evidence in favor of a close interaction between His189 and Tyr160 in the D2 polypeptide of photosystem II. This observation provides support for a model in which an imidazole nitrogen of His189 accepts the phenolic proton of Tyr160 upon oxidation of D, forming a back hydrogen bond to the phenolic oxygen of the neutral tyrosyl radical.

AB - The reaction center of photosystem II of oxygenic photosynthesis contains two redox-active tyrosines called Z and D, each of which can act as an electron donor to the oxidized primary electron donor, P680 +. These tyrosines are located in homologous positions on the third transmembrane α- helix of each of the two homologous polypeptides, D1 and D2, that comprise the reaction center. Tyrosine D of polypeptide D2 has been proposed, upon oxidation, to give up its phenolic proton to a nearby basic amino acid residue, forming a neutral radical. Modeling studies have pointed to His190 (spinach numbering) as a likely candidate for this basic residue. As a test of this hypothesis, we have constructed three site-directed mutations in the D2 polypeptide of the cyanobacterium Synechocystis sp. PCC6803. His189 (the Synechocystis homologue of His190 of spinach) has been replaced by glutamine, aspartate, or leucine. Instead of the normal D· EPR signal (g = 2.0046; line width 16-19 G), PSII core complexes isolated from these three mutants show an altered dark-stable EPR signal with a narrowed line width (11-13 G), and g values of 2.0046, 2.0043, and 2.0042 for the His189Gln, His189Asp, and His189Leu mutants, respectively. Despite the reduced line width, these EPR signals show g values and microwave-power saturation properties similar to the normal D· signal. Furthermore, specific deuteration in one of those mutants at the 3 and 5 positions of the phenol ring of the photosystem II reaction center tyrosines results in a loss of hyperfine structure of the EPR signal, proving that the signal indeed arises from tyrosine. Proton-ENDOR studies of these tyrosine radicals show that one hyperfine coupling component of 3.5-3.6 MHz, observed in the wild-type strain disappears in all three mutants. Upon incubation of wild-type photosystem II core complexes in D2O, this hyperfine coupling is lost, indicating that it originates from an exchangeable proton, most likely interacting with D· through a hydrogen bond. These results provide strong experimental evidence in favor of a close interaction between His189 and Tyr160 in the D2 polypeptide of photosystem II. This observation provides support for a model in which an imidazole nitrogen of His189 accepts the phenolic proton of Tyr160 upon oxidation of D, forming a back hydrogen bond to the phenolic oxygen of the neutral tyrosyl radical.

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U2 - 10.1021/bi00212a045

DO - 10.1021/bi00212a045

M3 - Article

VL - 32

SP - 13742

EP - 13748

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 49

ER -