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Title: Crystal structure of the pristine peroxidase ferryl center and its relevance to proton-coupled electron transfer

Abstract

The reaction of peroxides with peroxidases oxidizes the heme iron from Fe(III) to Fe(IV)=O and a porphyrin or aromatic side chain to a cationic radical. X-ray–generated hydrated electrons rapidly reduce Fe(IV), thereby requiring very short exposures using many crystals, and, even then, some reduction cannot be avoided. The new generation of X-ray free electron lasers capable of generating intense X-rays on the tenths of femtosecond time scale enables structure determination with no reduction or X-ray damage. Here, we report the 1.5-Å crystal structure of cytochrome c peroxidase (CCP) compound I (CmpI) using data obtained with the Stanford Linear Coherent Light Source (LCLS). This structure is consistent with previous structures. Of particular importance is the active site water structure that can mediate the proton transfer reactions required for both CmpI formation and reduction of Fe(IV)=O to Fe(III)-OH. The structures indicate that a water molecule is ideally positioned to shuttle protons between an iron-linked oxygen and the active site catalytic His. We therefore have carried out both computational and kinetic studies to probe the reduction of Fe(IV)=O. Kinetic solvent isotope experiments show that the transfer of a single proton is critical in the peroxidase rate-limiting step, which is very likely the proton-coupledmore » reduction of Fe(IV)=O to Fe(III)-OH. We also find that the pK a of the catalytic His substantially increases in CmpI, indicating that this active site His is the source of the proton required in the reduction of Fe(IV)=O to Fe(IV)-OH.« less

Authors:
; ; ; ; ; ; ; ;
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1236656
Grant/Contract Number:  
AC02-76SF00515
Resource Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 113 Journal Issue: 5; Journal ID: ISSN 0027-8424
Publisher:
Proceedings of the National Academy of Sciences
Country of Publication:
United States
Language:
English

Citation Formats

Chreifi, Georges, Baxter, Elizabeth L., Doukov, Tzanko, Cohen, Aina E., McPhillips, Scott E., Song, Jinhu, Meharenna, Yergalem T., Soltis, S. Michael, and Poulos, Thomas L. Crystal structure of the pristine peroxidase ferryl center and its relevance to proton-coupled electron transfer. United States: N. p., 2016. Web. doi:10.1073/pnas.1521664113.
Chreifi, Georges, Baxter, Elizabeth L., Doukov, Tzanko, Cohen, Aina E., McPhillips, Scott E., Song, Jinhu, Meharenna, Yergalem T., Soltis, S. Michael, & Poulos, Thomas L. Crystal structure of the pristine peroxidase ferryl center and its relevance to proton-coupled electron transfer. United States. doi:10.1073/pnas.1521664113.
Chreifi, Georges, Baxter, Elizabeth L., Doukov, Tzanko, Cohen, Aina E., McPhillips, Scott E., Song, Jinhu, Meharenna, Yergalem T., Soltis, S. Michael, and Poulos, Thomas L. Tue . "Crystal structure of the pristine peroxidase ferryl center and its relevance to proton-coupled electron transfer". United States. doi:10.1073/pnas.1521664113.
@article{osti_1236656,
title = {Crystal structure of the pristine peroxidase ferryl center and its relevance to proton-coupled electron transfer},
author = {Chreifi, Georges and Baxter, Elizabeth L. and Doukov, Tzanko and Cohen, Aina E. and McPhillips, Scott E. and Song, Jinhu and Meharenna, Yergalem T. and Soltis, S. Michael and Poulos, Thomas L.},
abstractNote = {The reaction of peroxides with peroxidases oxidizes the heme iron from Fe(III) to Fe(IV)=O and a porphyrin or aromatic side chain to a cationic radical. X-ray–generated hydrated electrons rapidly reduce Fe(IV), thereby requiring very short exposures using many crystals, and, even then, some reduction cannot be avoided. The new generation of X-ray free electron lasers capable of generating intense X-rays on the tenths of femtosecond time scale enables structure determination with no reduction or X-ray damage. Here, we report the 1.5-Å crystal structure of cytochrome c peroxidase (CCP) compound I (CmpI) using data obtained with the Stanford Linear Coherent Light Source (LCLS). This structure is consistent with previous structures. Of particular importance is the active site water structure that can mediate the proton transfer reactions required for both CmpI formation and reduction of Fe(IV)=O to Fe(III)-OH. The structures indicate that a water molecule is ideally positioned to shuttle protons between an iron-linked oxygen and the active site catalytic His. We therefore have carried out both computational and kinetic studies to probe the reduction of Fe(IV)=O. Kinetic solvent isotope experiments show that the transfer of a single proton is critical in the peroxidase rate-limiting step, which is very likely the proton-coupled reduction of Fe(IV)=O to Fe(III)-OH. We also find that the pK a of the catalytic His substantially increases in CmpI, indicating that this active site His is the source of the proton required in the reduction of Fe(IV)=O to Fe(IV)-OH.},
doi = {10.1073/pnas.1521664113},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 5,
volume = 113,
place = {United States},
year = {2016},
month = {1}
}

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DOI: 10.1073/pnas.1521664113

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Cited by: 13 works
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