FTIR spectroscopic evidence for the involvement of an acidic residue in quinone binding in cytochrome bd from Escherichia coli

Jie Zhang, Walter Oettmeier, Robert B. Gennis, Petra Hellwig

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

In this work, FTIR difference spectroscopy is used to search for possible binding partners and protonable groups involved in the binding of the quinol to cytochrome bd from Escherichia coli. In addition, the electrochemically induced FTIR difference spectra are compared for preparations of the enzyme isolated from cells grown at different oxygen levels in which the quinone content of the membrane is altered. On this basis, difference signals can be tentatively attributed to the vibrational modes of the different quinones types that are associated with the enzyme depending on growth conditions. Furthermore, vibrational modes due to the redox-dependent reorganization of the protein vary depending on the quinone associated with the isolated enzyme. Of particular interest are the observations that a mode at 1738 cm-1 is decreased and a mode at 1595 cm-1 is increased as observed in direct comparison to the data obtained from samples grown anaerobically. These signals indicate a change in the protonation state of an aspartic or glutamic acid. Since these changes are observed when the ubiquinone ratio in the preparation increases, the data provide evidence for the modulation of the binding site by the interacting quinone and the involvement of an acidic group in the binding site. The tentative assignments of the vibrational modes are supported by electrochemically induced FTIR difference spectra of cytochrome bd in the presence of the specific quinone binding site inhibitors heptylhydroxyquinoline-N-oxide (HQNO) or 2-methyl-3-undecylquinolone-4. Whereas HQNO leads to strong shifts in the FTIR redox difference spectrum, 2-methyl -3-undecylquinolone-4 induces a specific shift of a mode at 1635 cm-1, which likely originates from the displacement of the C=O group of the bound quinone.

Original languageEnglish (US)
Pages (from-to)4612-4617
Number of pages6
JournalBiochemistry
Volume41
Issue number14
DOIs
StatePublished - Apr 9 2002
Externally publishedYes

Fingerprint

Fourier Transform Infrared Spectroscopy
Cytochromes
Escherichia coli
Binding Sites
Oxidation-Reduction
Enzymes
Hydroquinones
Quinones
Ubiquinone
Protonation
Oxides
Glutamic Acid
Spectrum Analysis
Cells
Modulation
benzoquinone
Spectroscopy
Oxygen
Membranes
Growth

ASJC Scopus subject areas

  • Biochemistry

Cite this

FTIR spectroscopic evidence for the involvement of an acidic residue in quinone binding in cytochrome bd from Escherichia coli. / Zhang, Jie; Oettmeier, Walter; Gennis, Robert B.; Hellwig, Petra.

In: Biochemistry, Vol. 41, No. 14, 09.04.2002, p. 4612-4617.

Research output: Contribution to journalArticle

Zhang, Jie ; Oettmeier, Walter ; Gennis, Robert B. ; Hellwig, Petra. / FTIR spectroscopic evidence for the involvement of an acidic residue in quinone binding in cytochrome bd from Escherichia coli. In: Biochemistry. 2002 ; Vol. 41, No. 14. pp. 4612-4617.
@article{010735aeab5c47b784fc98d9ef2fc261,
title = "FTIR spectroscopic evidence for the involvement of an acidic residue in quinone binding in cytochrome bd from Escherichia coli",
abstract = "In this work, FTIR difference spectroscopy is used to search for possible binding partners and protonable groups involved in the binding of the quinol to cytochrome bd from Escherichia coli. In addition, the electrochemically induced FTIR difference spectra are compared for preparations of the enzyme isolated from cells grown at different oxygen levels in which the quinone content of the membrane is altered. On this basis, difference signals can be tentatively attributed to the vibrational modes of the different quinones types that are associated with the enzyme depending on growth conditions. Furthermore, vibrational modes due to the redox-dependent reorganization of the protein vary depending on the quinone associated with the isolated enzyme. Of particular interest are the observations that a mode at 1738 cm-1 is decreased and a mode at 1595 cm-1 is increased as observed in direct comparison to the data obtained from samples grown anaerobically. These signals indicate a change in the protonation state of an aspartic or glutamic acid. Since these changes are observed when the ubiquinone ratio in the preparation increases, the data provide evidence for the modulation of the binding site by the interacting quinone and the involvement of an acidic group in the binding site. The tentative assignments of the vibrational modes are supported by electrochemically induced FTIR difference spectra of cytochrome bd in the presence of the specific quinone binding site inhibitors heptylhydroxyquinoline-N-oxide (HQNO) or 2-methyl-3-undecylquinolone-4. Whereas HQNO leads to strong shifts in the FTIR redox difference spectrum, 2-methyl -3-undecylquinolone-4 induces a specific shift of a mode at 1635 cm-1, which likely originates from the displacement of the C=O group of the bound quinone.",
author = "Jie Zhang and Walter Oettmeier and Gennis, {Robert B.} and Petra Hellwig",
year = "2002",
month = "4",
day = "9",
doi = "10.1021/bi011784b",
language = "English (US)",
volume = "41",
pages = "4612--4617",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "14",

}

TY - JOUR

T1 - FTIR spectroscopic evidence for the involvement of an acidic residue in quinone binding in cytochrome bd from Escherichia coli

AU - Zhang, Jie

AU - Oettmeier, Walter

AU - Gennis, Robert B.

AU - Hellwig, Petra

PY - 2002/4/9

Y1 - 2002/4/9

N2 - In this work, FTIR difference spectroscopy is used to search for possible binding partners and protonable groups involved in the binding of the quinol to cytochrome bd from Escherichia coli. In addition, the electrochemically induced FTIR difference spectra are compared for preparations of the enzyme isolated from cells grown at different oxygen levels in which the quinone content of the membrane is altered. On this basis, difference signals can be tentatively attributed to the vibrational modes of the different quinones types that are associated with the enzyme depending on growth conditions. Furthermore, vibrational modes due to the redox-dependent reorganization of the protein vary depending on the quinone associated with the isolated enzyme. Of particular interest are the observations that a mode at 1738 cm-1 is decreased and a mode at 1595 cm-1 is increased as observed in direct comparison to the data obtained from samples grown anaerobically. These signals indicate a change in the protonation state of an aspartic or glutamic acid. Since these changes are observed when the ubiquinone ratio in the preparation increases, the data provide evidence for the modulation of the binding site by the interacting quinone and the involvement of an acidic group in the binding site. The tentative assignments of the vibrational modes are supported by electrochemically induced FTIR difference spectra of cytochrome bd in the presence of the specific quinone binding site inhibitors heptylhydroxyquinoline-N-oxide (HQNO) or 2-methyl-3-undecylquinolone-4. Whereas HQNO leads to strong shifts in the FTIR redox difference spectrum, 2-methyl -3-undecylquinolone-4 induces a specific shift of a mode at 1635 cm-1, which likely originates from the displacement of the C=O group of the bound quinone.

AB - In this work, FTIR difference spectroscopy is used to search for possible binding partners and protonable groups involved in the binding of the quinol to cytochrome bd from Escherichia coli. In addition, the electrochemically induced FTIR difference spectra are compared for preparations of the enzyme isolated from cells grown at different oxygen levels in which the quinone content of the membrane is altered. On this basis, difference signals can be tentatively attributed to the vibrational modes of the different quinones types that are associated with the enzyme depending on growth conditions. Furthermore, vibrational modes due to the redox-dependent reorganization of the protein vary depending on the quinone associated with the isolated enzyme. Of particular interest are the observations that a mode at 1738 cm-1 is decreased and a mode at 1595 cm-1 is increased as observed in direct comparison to the data obtained from samples grown anaerobically. These signals indicate a change in the protonation state of an aspartic or glutamic acid. Since these changes are observed when the ubiquinone ratio in the preparation increases, the data provide evidence for the modulation of the binding site by the interacting quinone and the involvement of an acidic group in the binding site. The tentative assignments of the vibrational modes are supported by electrochemically induced FTIR difference spectra of cytochrome bd in the presence of the specific quinone binding site inhibitors heptylhydroxyquinoline-N-oxide (HQNO) or 2-methyl-3-undecylquinolone-4. Whereas HQNO leads to strong shifts in the FTIR redox difference spectrum, 2-methyl -3-undecylquinolone-4 induces a specific shift of a mode at 1635 cm-1, which likely originates from the displacement of the C=O group of the bound quinone.

UR - http://www.scopus.com/inward/record.url?scp=0037046156&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037046156&partnerID=8YFLogxK

U2 - 10.1021/bi011784b

DO - 10.1021/bi011784b

M3 - Article

VL - 41

SP - 4612

EP - 4617

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 14

ER -