Protein Oxidation in the Lungs of C57BL/6J Mice Following X-Irradiation
Damage to normal lung tissue is a limiting factor when ionizing radiation is used in clinical applications. In addition, radiation pneumonitis and fibrosis are a major cause of mortality following accidental radiation exposure in humans. Although clinical symptoms may not develop for months after ra...
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| Format: | Article |
| Language: | English |
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MDPI AG
2015-08-01
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| Series: | Proteomes |
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| Online Access: | http://www.mdpi.com/2227-7382/3/3/249 |
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| author | Michal Barshishat-Kupper Elizabeth A. McCart James G. Freedy Ashlee J. Tipton Vitaly Nagy Sung-Yop Kim Michael R. Landauer Gregory P. Mueller Regina M. Day |
| author_facet | Michal Barshishat-Kupper Elizabeth A. McCart James G. Freedy Ashlee J. Tipton Vitaly Nagy Sung-Yop Kim Michael R. Landauer Gregory P. Mueller Regina M. Day |
| author_sort | Michal Barshishat-Kupper |
| collection | DOAJ |
| description | Damage to normal lung tissue is a limiting factor when ionizing radiation is used in clinical applications. In addition, radiation pneumonitis and fibrosis are a major cause of mortality following accidental radiation exposure in humans. Although clinical symptoms may not develop for months after radiation exposure, immediate events induced by radiation are believed to generate molecular and cellular cascades that proceed during a clinical latent period. Oxidative damage to DNA is considered a primary cause of radiation injury to cells. DNA can be repaired by highly efficient mechanisms while repair of oxidized proteins is limited. Oxidized proteins are often destined for degradation. We examined protein oxidation following 17 Gy (0.6 Gy/min) thoracic X-irradiation in C57BL/6J mice. Seventeen Gy thoracic irradiation resulted in 100% mortality of mice within 127–189 days postirradiation. Necropsy findings indicated that pneumonitis and pulmonary fibrosis were the leading cause of mortality. We investigated the oxidation of lung proteins at 24 h postirradiation following 17 Gy thoracic irradiation using 2-D gel electrophoresis and OxyBlot for the detection of protein carbonylation. Seven carbonylated proteins were identified using mass spectrometry: serum albumin, selenium binding protein-1, alpha antitrypsin, cytoplasmic actin-1, carbonic anhydrase-2, peroxiredoxin-6, and apolipoprotein A1. The carbonylation status of carbonic anhydrase-2, selenium binding protein, and peroxiredoxin-6 was higher in control lung tissue. Apolipoprotein A1 and serum albumin carbonylation were increased following X-irradiation, as confirmed by OxyBlot immunoprecipitation and Western blotting. Our findings indicate that the profile of specific protein oxidation in the lung is altered following radiation exposure. |
| first_indexed | 2024-04-14T02:00:41Z |
| format | Article |
| id | doaj.art-ddead878dff945efb568cb7b017775ac |
| institution | Directory Open Access Journal |
| issn | 2227-7382 |
| language | English |
| last_indexed | 2024-04-14T02:00:41Z |
| publishDate | 2015-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Proteomes |
| spelling | doaj.art-ddead878dff945efb568cb7b017775ac2022-12-22T02:18:50ZengMDPI AGProteomes2227-73822015-08-013324926510.3390/proteomes3030249proteomes3030249Protein Oxidation in the Lungs of C57BL/6J Mice Following X-IrradiationMichal Barshishat-Kupper0Elizabeth A. McCart1James G. Freedy2Ashlee J. Tipton3Vitaly Nagy4Sung-Yop Kim5Michael R. Landauer6Gregory P. Mueller7Regina M. Day8Department of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USADepartment of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USADepartment of Anatomy, Physiology and Genetics, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USADepartment of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USAOperational Dosimetry Division, Armed Forces Radiobiology Research Institute, Uniformed Services University of the Health Sciences, Bethesda, MD 20889, USAOperational Dosimetry Division, Armed Forces Radiobiology Research Institute, Uniformed Services University of the Health Sciences, Bethesda, MD 20889, USARadiation Countermeasures Program, Scientific Research Department, Armed Forces Radiobiology Research Institute, Uniformed Services University of the Health Sciences, Bethesda, MD 20889, USADepartment of Anatomy, Physiology and Genetics, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USADepartment of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USADamage to normal lung tissue is a limiting factor when ionizing radiation is used in clinical applications. In addition, radiation pneumonitis and fibrosis are a major cause of mortality following accidental radiation exposure in humans. Although clinical symptoms may not develop for months after radiation exposure, immediate events induced by radiation are believed to generate molecular and cellular cascades that proceed during a clinical latent period. Oxidative damage to DNA is considered a primary cause of radiation injury to cells. DNA can be repaired by highly efficient mechanisms while repair of oxidized proteins is limited. Oxidized proteins are often destined for degradation. We examined protein oxidation following 17 Gy (0.6 Gy/min) thoracic X-irradiation in C57BL/6J mice. Seventeen Gy thoracic irradiation resulted in 100% mortality of mice within 127–189 days postirradiation. Necropsy findings indicated that pneumonitis and pulmonary fibrosis were the leading cause of mortality. We investigated the oxidation of lung proteins at 24 h postirradiation following 17 Gy thoracic irradiation using 2-D gel electrophoresis and OxyBlot for the detection of protein carbonylation. Seven carbonylated proteins were identified using mass spectrometry: serum albumin, selenium binding protein-1, alpha antitrypsin, cytoplasmic actin-1, carbonic anhydrase-2, peroxiredoxin-6, and apolipoprotein A1. The carbonylation status of carbonic anhydrase-2, selenium binding protein, and peroxiredoxin-6 was higher in control lung tissue. Apolipoprotein A1 and serum albumin carbonylation were increased following X-irradiation, as confirmed by OxyBlot immunoprecipitation and Western blotting. Our findings indicate that the profile of specific protein oxidation in the lung is altered following radiation exposure.http://www.mdpi.com/2227-7382/3/3/249thoracic irradiationprotein carbonylationOxyBlotmass spectrometry2-D gel electrophoresispulmonary fibrosisradiation pneumonitis |
| spellingShingle | Michal Barshishat-Kupper Elizabeth A. McCart James G. Freedy Ashlee J. Tipton Vitaly Nagy Sung-Yop Kim Michael R. Landauer Gregory P. Mueller Regina M. Day Protein Oxidation in the Lungs of C57BL/6J Mice Following X-Irradiation Proteomes thoracic irradiation protein carbonylation OxyBlot mass spectrometry 2-D gel electrophoresis pulmonary fibrosis radiation pneumonitis |
| title | Protein Oxidation in the Lungs of C57BL/6J Mice Following X-Irradiation |
| title_full | Protein Oxidation in the Lungs of C57BL/6J Mice Following X-Irradiation |
| title_fullStr | Protein Oxidation in the Lungs of C57BL/6J Mice Following X-Irradiation |
| title_full_unstemmed | Protein Oxidation in the Lungs of C57BL/6J Mice Following X-Irradiation |
| title_short | Protein Oxidation in the Lungs of C57BL/6J Mice Following X-Irradiation |
| title_sort | protein oxidation in the lungs of c57bl 6j mice following x irradiation |
| topic | thoracic irradiation protein carbonylation OxyBlot mass spectrometry 2-D gel electrophoresis pulmonary fibrosis radiation pneumonitis |
| url | http://www.mdpi.com/2227-7382/3/3/249 |
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