Oxidation of myosin by haem proteins generates myosin radicals and protein cross-links

Marianne Lund Lametsch; Catherine Luxford; Leif Horsfelt Skibsted; Michael Jonathan Davies

doi:10.1042/BJ20071107

Oxidation of myosin by haem proteins generates myosin radicals and protein cross-links

Marianne Lund Lametsch, Catherine Luxford, Leif Horsfelt Skibsted, Michael Jonathan Davies

87 Citations (Scopus)

Abstract

Previous studies have reported that myosin can be modified by oxidative stress and particularly by activated haem proteins. These reactions have been implicated in changes in the properties of this protein in food samples (changes in meat tenderness and palatability), in human physiology (alteration of myocyte function and force generation) and in disease (e.g. cardiomyopathy, chronic heart failure). The oxidant species, mechanisms of reaction and consequences of these reactions are incompletely characterized. In the present study, the nature of the transient species generated on myosin as a result of the reaction with activated haem proteins (horseradish peroxidase/H2O2) and met-myoglobin/H2O2) has been investigated by EPR spectroscopy and amino-acid consumption, product formation has been characterized by HPLC, and changes in protein integrity have been determined by SDS/PAGE. Multiple radical species have been detected by EPR in both the presence and the absence of spin traps. Evidence has been obtained for the presence of thiyl, tyrosyl and other unidentified radical species on myosin as a result of damage-transfer from oxidized myoglobin or horseradish peroxidase. The generation of thiyl and tyrosyl radicals is consistent with the observed consumption of cysteine and tyrosine residues, the detection of di-tyrosine by HPLC and the detection of both reducible (disulfide bond) and non-reducible cross-links between myosin molecules by SDS/PAGE. The time course of radical formation on myosin, product generation and cross-link induction are consistent with these processes being interlinked. These changes are consistent with the altered function and properties of myosin in muscle tissue exposed to oxidative stress arising from disease or from food processing.

Original language	English
Journal	Biochemical Journal
Volume	410
Issue number	3
Pages (from-to)	565-574
Number of pages	10
ISSN	0264-6021
DOIs	https://doi.org/10.1042/BJ20071107
Publication status	Published - 2008

Keywords

Amino Acids
Animals
Chromatography, High Pressure Liquid
Electron Spin Resonance Spectroscopy
Electrophoresis, Polyacrylamide Gel
Hemeproteins
Myosins
Oxidation-Reduction
Protein Conformation
Spin Labels
Swine

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1042/BJ20071107

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title = "Oxidation of myosin by haem proteins generates myosin radicals and protein cross-links",

abstract = "Previous studies have reported that myosin can be modified by oxidative stress and particularly by activated haem proteins. These reactions have been implicated in changes in the properties of this protein in food samples (changes in meat tenderness and palatability), in human physiology (alteration of myocyte function and force generation) and in disease (e.g. cardiomyopathy, chronic heart failure). The oxidant species, mechanisms of reaction and consequences of these reactions are incompletely characterized. In the present study, the nature of the transient species generated on myosin as a result of the reaction with activated haem proteins (horseradish peroxidase/H2O2) and met-myoglobin/H2O2) has been investigated by EPR spectroscopy and amino-acid consumption, product formation has been characterized by HPLC, and changes in protein integrity have been determined by SDS/PAGE. Multiple radical species have been detected by EPR in both the presence and the absence of spin traps. Evidence has been obtained for the presence of thiyl, tyrosyl and other unidentified radical species on myosin as a result of damage-transfer from oxidized myoglobin or horseradish peroxidase. The generation of thiyl and tyrosyl radicals is consistent with the observed consumption of cysteine and tyrosine residues, the detection of di-tyrosine by HPLC and the detection of both reducible (disulfide bond) and non-reducible cross-links between myosin molecules by SDS/PAGE. The time course of radical formation on myosin, product generation and cross-link induction are consistent with these processes being interlinked. These changes are consistent with the altered function and properties of myosin in muscle tissue exposed to oxidative stress arising from disease or from food processing.",

keywords = "Amino Acids, Animals, Chromatography, High Pressure Liquid, Electron Spin Resonance Spectroscopy, Electrophoresis, Polyacrylamide Gel, Hemeproteins, Myosins, Oxidation-Reduction, Protein Conformation, Spin Labels, Swine",

author = "Lametsch, {Marianne Lund} and Catherine Luxford and Skibsted, {Leif Horsfelt} and Davies, {Michael Jonathan}",

year = "2008",

doi = "10.1042/BJ20071107",

language = "English",

volume = "410",

pages = "565--574",

journal = "Biochemical Journal",

issn = "0264-6021",

publisher = "Portland Press Ltd.",

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TY - JOUR

T1 - Oxidation of myosin by haem proteins generates myosin radicals and protein cross-links

AU - Lametsch, Marianne Lund

AU - Luxford, Catherine

AU - Skibsted, Leif Horsfelt

AU - Davies, Michael Jonathan

PY - 2008

Y1 - 2008

N2 - Previous studies have reported that myosin can be modified by oxidative stress and particularly by activated haem proteins. These reactions have been implicated in changes in the properties of this protein in food samples (changes in meat tenderness and palatability), in human physiology (alteration of myocyte function and force generation) and in disease (e.g. cardiomyopathy, chronic heart failure). The oxidant species, mechanisms of reaction and consequences of these reactions are incompletely characterized. In the present study, the nature of the transient species generated on myosin as a result of the reaction with activated haem proteins (horseradish peroxidase/H2O2) and met-myoglobin/H2O2) has been investigated by EPR spectroscopy and amino-acid consumption, product formation has been characterized by HPLC, and changes in protein integrity have been determined by SDS/PAGE. Multiple radical species have been detected by EPR in both the presence and the absence of spin traps. Evidence has been obtained for the presence of thiyl, tyrosyl and other unidentified radical species on myosin as a result of damage-transfer from oxidized myoglobin or horseradish peroxidase. The generation of thiyl and tyrosyl radicals is consistent with the observed consumption of cysteine and tyrosine residues, the detection of di-tyrosine by HPLC and the detection of both reducible (disulfide bond) and non-reducible cross-links between myosin molecules by SDS/PAGE. The time course of radical formation on myosin, product generation and cross-link induction are consistent with these processes being interlinked. These changes are consistent with the altered function and properties of myosin in muscle tissue exposed to oxidative stress arising from disease or from food processing.

AB - Previous studies have reported that myosin can be modified by oxidative stress and particularly by activated haem proteins. These reactions have been implicated in changes in the properties of this protein in food samples (changes in meat tenderness and palatability), in human physiology (alteration of myocyte function and force generation) and in disease (e.g. cardiomyopathy, chronic heart failure). The oxidant species, mechanisms of reaction and consequences of these reactions are incompletely characterized. In the present study, the nature of the transient species generated on myosin as a result of the reaction with activated haem proteins (horseradish peroxidase/H2O2) and met-myoglobin/H2O2) has been investigated by EPR spectroscopy and amino-acid consumption, product formation has been characterized by HPLC, and changes in protein integrity have been determined by SDS/PAGE. Multiple radical species have been detected by EPR in both the presence and the absence of spin traps. Evidence has been obtained for the presence of thiyl, tyrosyl and other unidentified radical species on myosin as a result of damage-transfer from oxidized myoglobin or horseradish peroxidase. The generation of thiyl and tyrosyl radicals is consistent with the observed consumption of cysteine and tyrosine residues, the detection of di-tyrosine by HPLC and the detection of both reducible (disulfide bond) and non-reducible cross-links between myosin molecules by SDS/PAGE. The time course of radical formation on myosin, product generation and cross-link induction are consistent with these processes being interlinked. These changes are consistent with the altered function and properties of myosin in muscle tissue exposed to oxidative stress arising from disease or from food processing.

KW - Amino Acids

KW - Animals

KW - Chromatography, High Pressure Liquid

KW - Electron Spin Resonance Spectroscopy

KW - Electrophoresis, Polyacrylamide Gel

KW - Hemeproteins

KW - Myosins

KW - Oxidation-Reduction

KW - Protein Conformation

KW - Spin Labels

KW - Swine

U2 - 10.1042/BJ20071107

DO - 10.1042/BJ20071107

M3 - Journal article

C2 - 18039181

SN - 0264-6021

VL - 410

SP - 565

EP - 574

JO - Biochemical Journal

JF - Biochemical Journal

IS - 3

ER -

Oxidation of myosin by haem proteins generates myosin radicals and protein cross-links

Abstract

Keywords

UN SDGs

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