Cytochrome c peroxidase complexed with cytochrome c has an unperturbed heme moiety

Jianling Wang, Randy W. Larsen, Susan J. Moench, James D. Satterlee, Denis L. Rousseau, Mark R. Ondrias

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Transient resonance Raman, Raman difference, circular dichroism (CD), and optical absorption studies have been carried out on the electrostatic complexes formed by yeast cytochrome c peroxidase (CCP) with horse cytochrome c (Cytc) in low ionic strength solutions. In all the complexes examined [e.g., CCP(II)/Cytc(II), CCP(III)/Cytc(II), CCP(III)/Cytc(III)], the local heme environments of both proteins are largely unperturbed upon complexation. Specifically, CCP preserves a completely pentacoordinate high-spin home in both its ferric and ferrous forms in CCP/Cytc complexes and uncomplexed mixtures. We found no evidence corroborating the previously reported increase in the low-spin fraction of CCP heme upon complexation with Cytc [Hildebrandt et al. (1992) Biochemistry 31, 2384-2392]. Instead, our Raman data strongly suggest that the H-bonding networks in the distal and proximal pockets of CCP are well maintained in the complexes. On the other hand, CD spectra of CCP(III)/Cytc(III) complexes showed substantial variations (relative to the uncomplexed mixtures) in the far-UV region, reflecting some protein conformational rearrangements. In addition, the spectral data suggest that complexation with Cytc affects the previously observed pH-dependent flexibility of the heme structure of CCP and thus influences the photodynamics of the CCP active site.

Original languageEnglish (US)
Pages (from-to)453-463
Number of pages11
JournalBiochemistry
Volume35
Issue number2
DOIs
StatePublished - Jan 16 1996
Externally publishedYes

Fingerprint

Cytochrome-c Peroxidase
Cytochromes c
Heme
Complexation
Circular Dichroism
Biochemistry
Ionic strength
Complex Mixtures
Static Electricity
Yeast
Osmolar Concentration
Light absorption
Horses
Electrostatics
Catalytic Domain

ASJC Scopus subject areas

  • Biochemistry

Cite this

Wang, J., Larsen, R. W., Moench, S. J., Satterlee, J. D., Rousseau, D. L., & Ondrias, M. R. (1996). Cytochrome c peroxidase complexed with cytochrome c has an unperturbed heme moiety. Biochemistry, 35(2), 453-463. https://doi.org/10.1021/bi9518499

Cytochrome c peroxidase complexed with cytochrome c has an unperturbed heme moiety. / Wang, Jianling; Larsen, Randy W.; Moench, Susan J.; Satterlee, James D.; Rousseau, Denis L.; Ondrias, Mark R.

In: Biochemistry, Vol. 35, No. 2, 16.01.1996, p. 453-463.

Research output: Contribution to journalArticle

Wang, J, Larsen, RW, Moench, SJ, Satterlee, JD, Rousseau, DL & Ondrias, MR 1996, 'Cytochrome c peroxidase complexed with cytochrome c has an unperturbed heme moiety', Biochemistry, vol. 35, no. 2, pp. 453-463. https://doi.org/10.1021/bi9518499
Wang, Jianling ; Larsen, Randy W. ; Moench, Susan J. ; Satterlee, James D. ; Rousseau, Denis L. ; Ondrias, Mark R. / Cytochrome c peroxidase complexed with cytochrome c has an unperturbed heme moiety. In: Biochemistry. 1996 ; Vol. 35, No. 2. pp. 453-463.
@article{a9d606d9ba6d42e7be8817879b5bbe0f,
title = "Cytochrome c peroxidase complexed with cytochrome c has an unperturbed heme moiety",
abstract = "Transient resonance Raman, Raman difference, circular dichroism (CD), and optical absorption studies have been carried out on the electrostatic complexes formed by yeast cytochrome c peroxidase (CCP) with horse cytochrome c (Cytc) in low ionic strength solutions. In all the complexes examined [e.g., CCP(II)/Cytc(II), CCP(III)/Cytc(II), CCP(III)/Cytc(III)], the local heme environments of both proteins are largely unperturbed upon complexation. Specifically, CCP preserves a completely pentacoordinate high-spin home in both its ferric and ferrous forms in CCP/Cytc complexes and uncomplexed mixtures. We found no evidence corroborating the previously reported increase in the low-spin fraction of CCP heme upon complexation with Cytc [Hildebrandt et al. (1992) Biochemistry 31, 2384-2392]. Instead, our Raman data strongly suggest that the H-bonding networks in the distal and proximal pockets of CCP are well maintained in the complexes. On the other hand, CD spectra of CCP(III)/Cytc(III) complexes showed substantial variations (relative to the uncomplexed mixtures) in the far-UV region, reflecting some protein conformational rearrangements. In addition, the spectral data suggest that complexation with Cytc affects the previously observed pH-dependent flexibility of the heme structure of CCP and thus influences the photodynamics of the CCP active site.",
author = "Jianling Wang and Larsen, {Randy W.} and Moench, {Susan J.} and Satterlee, {James D.} and Rousseau, {Denis L.} and Ondrias, {Mark R.}",
year = "1996",
month = "1",
day = "16",
doi = "10.1021/bi9518499",
language = "English (US)",
volume = "35",
pages = "453--463",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "2",

}

TY - JOUR

T1 - Cytochrome c peroxidase complexed with cytochrome c has an unperturbed heme moiety

AU - Wang, Jianling

AU - Larsen, Randy W.

AU - Moench, Susan J.

AU - Satterlee, James D.

AU - Rousseau, Denis L.

AU - Ondrias, Mark R.

PY - 1996/1/16

Y1 - 1996/1/16

N2 - Transient resonance Raman, Raman difference, circular dichroism (CD), and optical absorption studies have been carried out on the electrostatic complexes formed by yeast cytochrome c peroxidase (CCP) with horse cytochrome c (Cytc) in low ionic strength solutions. In all the complexes examined [e.g., CCP(II)/Cytc(II), CCP(III)/Cytc(II), CCP(III)/Cytc(III)], the local heme environments of both proteins are largely unperturbed upon complexation. Specifically, CCP preserves a completely pentacoordinate high-spin home in both its ferric and ferrous forms in CCP/Cytc complexes and uncomplexed mixtures. We found no evidence corroborating the previously reported increase in the low-spin fraction of CCP heme upon complexation with Cytc [Hildebrandt et al. (1992) Biochemistry 31, 2384-2392]. Instead, our Raman data strongly suggest that the H-bonding networks in the distal and proximal pockets of CCP are well maintained in the complexes. On the other hand, CD spectra of CCP(III)/Cytc(III) complexes showed substantial variations (relative to the uncomplexed mixtures) in the far-UV region, reflecting some protein conformational rearrangements. In addition, the spectral data suggest that complexation with Cytc affects the previously observed pH-dependent flexibility of the heme structure of CCP and thus influences the photodynamics of the CCP active site.

AB - Transient resonance Raman, Raman difference, circular dichroism (CD), and optical absorption studies have been carried out on the electrostatic complexes formed by yeast cytochrome c peroxidase (CCP) with horse cytochrome c (Cytc) in low ionic strength solutions. In all the complexes examined [e.g., CCP(II)/Cytc(II), CCP(III)/Cytc(II), CCP(III)/Cytc(III)], the local heme environments of both proteins are largely unperturbed upon complexation. Specifically, CCP preserves a completely pentacoordinate high-spin home in both its ferric and ferrous forms in CCP/Cytc complexes and uncomplexed mixtures. We found no evidence corroborating the previously reported increase in the low-spin fraction of CCP heme upon complexation with Cytc [Hildebrandt et al. (1992) Biochemistry 31, 2384-2392]. Instead, our Raman data strongly suggest that the H-bonding networks in the distal and proximal pockets of CCP are well maintained in the complexes. On the other hand, CD spectra of CCP(III)/Cytc(III) complexes showed substantial variations (relative to the uncomplexed mixtures) in the far-UV region, reflecting some protein conformational rearrangements. In addition, the spectral data suggest that complexation with Cytc affects the previously observed pH-dependent flexibility of the heme structure of CCP and thus influences the photodynamics of the CCP active site.

UR - http://www.scopus.com/inward/record.url?scp=0030053508&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030053508&partnerID=8YFLogxK

U2 - 10.1021/bi9518499

DO - 10.1021/bi9518499

M3 - Article

VL - 35

SP - 453

EP - 463

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 2

ER -