Accumulation of isolevuglandin-modified protein in normal and fibrotic lung.
Protein lysine modification by γ-ketoaldehyde isomers derived from arachidonic acid, termed isolevuglandins (IsoLGs), is emerging as a mechanistic link between pathogenic reactive oxygen species and disease progression. However, the questions of whether covalent modification of proteins by IsoLGs are subject to genetic regulation and the identity of IsoLG-modified proteins remain unclear. Herein we show that Nrf2 and Nox2 are key regulators of IsoLG modification in pulmonary tissue and report on the identity of proteins analyzed by LC-MS following immunoaffinity purification of IsoLG-modified proteins. Gene ontology analysis revealed that proteins in numerous cellular pathways are susceptible to IsoLG modification. Although cells tolerate basal levels of modification, exceeding them induces apoptosis. We found prominent modification in a murine model of radiation-induced pulmonary fibrosis and in idiopathic pulmonary fibrosis, two diseases considered to be promoted by gene-regulated oxidant stress. Based on these results we hypothesize that IsoLG modification is a hitherto unrecognized sequelae that contributes to radiation-induced pulmonary injury and IPF.
Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, TN 37240, USA.|Department of Radiation Oncology, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
Stacey
S
Mont
Division of Clinical Pharmacology, Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
Sean S
SS
Davies
Division of Clinical Pharmacology, Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
L Jackson
LJ
Roberts Second
Department of Biochemistry, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
Raymond L
RL
Mernaugh
Department of Biochemistry, Vanderbilt University Medical Center, Nashville, TN 37240, USA.|Proteomics Laboratory and Mass Spectrometry Research Center, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
W Hayes
WH
McDonald
Department of Medicine, Department of Immunology, Roswell Park Cancer Institute, and University at Buffalo Jacobs School of Medicine and Biomedical Sciences, Buffalo, NY, 14263, USA.
Brahm H
BH
Segal
Division of Clinical Pharmacology, Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
William
W
Zackert
Division of Pulmonary &Critical Care, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
Jonathan A
JA
Kropski
Division of Pulmonary &Critical Care, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
Timothy S
TS
Blackwell
Department of Radiation Oncology, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
Konjeti R
KR
Sekhar
Department of Biochemistry, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
James J
JJ
Galligan
Division of Pulmonary &Critical Care, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
Pierre P
PP
Massion
Department of Biochemistry, Vanderbilt University Medical Center, Nashville, TN 37240, USA.|A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Vanderbilt Institute of Chemical Biology, Vanderbilt-Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
Lawrence J
LJ
Marnett
Department of Experimental Radiation Oncology, Division of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77230, USA.
Elizabeth L
EL
Travis
Department of Radiation Oncology, Vanderbilt University Medical Center, Nashville, TN 37240, USA.
Michael L
ML
Freeman
D000818:Animals; D002853:Chromatography, Liquid; D004195:Disease Models, Animal; D008168:Lung / Q000737:chemistry* / Q000378:metabolism*; D013058:Mass Spectrometry; D051379:Mice; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis*; D011658:Pulmonary Fibrosis / Q000473:pathology*
D020543:Proteome