Determination by X-ray absorption spectroscopy of the Fe-Fe separation in the oxidized form of the hydrxylase of methane monooxygenase alone and in the presence of MMOD

D.J. Rudd, M.H. Sazinsky, M. Merkx, S.J. Lippard, B. Hedman, K.O. Hodgson

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Abstract

The diiron active site in the hydroxylase of Methylococcus capsulatus (Bath) methane monooxygenase (MMOH) has been studied in the oxidized form by X-ray absorption spectroscopy (XAS). Previous investigations by XAS and X-ray crystallography have identified two different distances (3.0 and 3.4 Å) between the two Fe atoms in the dinuclear site. The present study has employed a systematic extended X-ray absorption fine structure (EXAFS) fitting methodology, utilizing known and simulated active site and relevant model structures, to determine unambiguously the Fe-Fe separation in the oxidized form of MMOH. Consistent and unique fits were only possible for an Fe-Fe distance of 3.0 Å. This methodology was then applied to study potential changes in the active site local structure in the presence of MMOD, a protein of unknown function in multicomponent MMO. Fe K-edge and EXAFS analyses revealed negligible changes in the diiron site electronic and geometric structure upon addition of MMOD to oxidized MMOH.
Original languageEnglish
Pages (from-to)4579-4589
JournalInorganic Chemistry
Volume43
Issue number15
DOIs
Publication statusPublished - 2004

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methane monooxygenase
X ray absorption spectroscopy
X ray absorption
absorption spectroscopy
methane
X ray crystallography
Model structures
Mixed Function Oxygenases
x rays
Atoms
fine structure
methodology
Proteins
crystallography
baths
electronic structure
proteins

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@article{80f883891fac4de5a33443bcc84c51fb,
title = "Determination by X-ray absorption spectroscopy of the Fe-Fe separation in the oxidized form of the hydrxylase of methane monooxygenase alone and in the presence of MMOD",
abstract = "The diiron active site in the hydroxylase of Methylococcus capsulatus (Bath) methane monooxygenase (MMOH) has been studied in the oxidized form by X-ray absorption spectroscopy (XAS). Previous investigations by XAS and X-ray crystallography have identified two different distances (3.0 and 3.4 {\AA}) between the two Fe atoms in the dinuclear site. The present study has employed a systematic extended X-ray absorption fine structure (EXAFS) fitting methodology, utilizing known and simulated active site and relevant model structures, to determine unambiguously the Fe-Fe separation in the oxidized form of MMOH. Consistent and unique fits were only possible for an Fe-Fe distance of 3.0 {\AA}. This methodology was then applied to study potential changes in the active site local structure in the presence of MMOD, a protein of unknown function in multicomponent MMO. Fe K-edge and EXAFS analyses revealed negligible changes in the diiron site electronic and geometric structure upon addition of MMOD to oxidized MMOH.",
author = "D.J. Rudd and M.H. Sazinsky and M. Merkx and S.J. Lippard and B. Hedman and K.O. Hodgson",
year = "2004",
doi = "10.1021/ic049716b",
language = "English",
volume = "43",
pages = "4579--4589",
journal = "Inorganic Chemistry",
issn = "0020-1669",
publisher = "American Chemical Society",
number = "15",

}

Determination by X-ray absorption spectroscopy of the Fe-Fe separation in the oxidized form of the hydrxylase of methane monooxygenase alone and in the presence of MMOD. / Rudd, D.J.; Sazinsky, M.H.; Merkx, M.; Lippard, S.J.; Hedman, B.; Hodgson, K.O.

In: Inorganic Chemistry, Vol. 43, No. 15, 2004, p. 4579-4589.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Determination by X-ray absorption spectroscopy of the Fe-Fe separation in the oxidized form of the hydrxylase of methane monooxygenase alone and in the presence of MMOD

AU - Rudd, D.J.

AU - Sazinsky, M.H.

AU - Merkx, M.

AU - Lippard, S.J.

AU - Hedman, B.

AU - Hodgson, K.O.

PY - 2004

Y1 - 2004

N2 - The diiron active site in the hydroxylase of Methylococcus capsulatus (Bath) methane monooxygenase (MMOH) has been studied in the oxidized form by X-ray absorption spectroscopy (XAS). Previous investigations by XAS and X-ray crystallography have identified two different distances (3.0 and 3.4 Å) between the two Fe atoms in the dinuclear site. The present study has employed a systematic extended X-ray absorption fine structure (EXAFS) fitting methodology, utilizing known and simulated active site and relevant model structures, to determine unambiguously the Fe-Fe separation in the oxidized form of MMOH. Consistent and unique fits were only possible for an Fe-Fe distance of 3.0 Å. This methodology was then applied to study potential changes in the active site local structure in the presence of MMOD, a protein of unknown function in multicomponent MMO. Fe K-edge and EXAFS analyses revealed negligible changes in the diiron site electronic and geometric structure upon addition of MMOD to oxidized MMOH.

AB - The diiron active site in the hydroxylase of Methylococcus capsulatus (Bath) methane monooxygenase (MMOH) has been studied in the oxidized form by X-ray absorption spectroscopy (XAS). Previous investigations by XAS and X-ray crystallography have identified two different distances (3.0 and 3.4 Å) between the two Fe atoms in the dinuclear site. The present study has employed a systematic extended X-ray absorption fine structure (EXAFS) fitting methodology, utilizing known and simulated active site and relevant model structures, to determine unambiguously the Fe-Fe separation in the oxidized form of MMOH. Consistent and unique fits were only possible for an Fe-Fe distance of 3.0 Å. This methodology was then applied to study potential changes in the active site local structure in the presence of MMOD, a protein of unknown function in multicomponent MMO. Fe K-edge and EXAFS analyses revealed negligible changes in the diiron site electronic and geometric structure upon addition of MMOD to oxidized MMOH.

U2 - 10.1021/ic049716b

DO - 10.1021/ic049716b

M3 - Article

VL - 43

SP - 4579

EP - 4589

JO - Inorganic Chemistry

JF - Inorganic Chemistry

SN - 0020-1669

IS - 15

ER -