High resolution mass spectrometric alveolar proteomics: Identification of surfactant protein SP‐A and SP‐D modifications in proteinosis and cystic fibrosis patients

Bai, Yu; Galetskiy, Dmitry; Damoc, Eugen; Paschen, Christian; Liu, Zhiqiang; Griese, Mathias; Liu, Shuying; Przybylski, Michael

doi:10.1002/pmic.200400855

Cited by 41 publications

(42 citation statements)

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“…Proteolytic derivatives of surfactant protein A (SP-A), an important innate host defense component of the lungs, was found present in BALF of cystic fibrosis (CF) patients but not present in controls [415]. Using a high-resolution FT-ICR MS analysis, surfactant proteins SP-A and SP-D, including structurally modified and truncated forms, were identified in BALF from patients with CF, chronic bronchitis, and pulmonary alveolar proteinosis [406].…”

Section: Application To Human Diseases Detectionmentioning

confidence: 99%

“…Monitoring the proteolytic changes and structurally modified pattern of BALF proteins in lung diseases may provide further insight of the disease mechanism or diagnostic potential [397,406,415,416]. Most lung disorders are known to be associated to considerable modifications of surfactant composition.…”

Section: Application To Human Diseases Detectionmentioning

confidence: 99%

“…The current master gel of BALF proteins comprises more than 1200 spots visualized by silver staining [402]. Identification of proteins on 2-D gels was initially performed using immunoblotting, Edman sequencing, or matching to a reference gel [403,404], and this was significantly improved through the use of advanced MS identification techniques [405][406][407][408][409]. A large-scale proteomic analysis by using immunoaffinity depletion, SDS-PAGE fractionation, protein in-gel digestion, and subsequent nano-LC-MS/MS was reported for identification and semiquantitation of more than 1500 distinct proteins in BALF.…”

Section: Analysis and Characterization Of Balf Proteomementioning

confidence: 99%

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Human body fluid proteome analysis

2006

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The focus of this article is to review the recent advances in proteome analysis of human body fluids, including plasma/serum, urine, cerebrospinal fluid, saliva, bronchoalveolar lavage fluid, synovial fluid, nipple aspirate fluid, tear fluid, and amniotic fluid, as well as its applications to human disease biomarker discovery. We aim to summarize the proteomics technologies currently used for global identification and quantification of body fluid proteins, and elaborate the putative biomarkers discovered for a variety of human diseases through human body fluid proteome (HBFP) analysis. Some critical concerns and perspectives in this emerging field are also discussed. With the advances made in proteomics technologies, the impact of HBFP analysis in the search for clinically relevant disease biomarkers would be realized in the future.

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Section: Application To Human Diseases Detectionmentioning

confidence: 99%

Section: Application To Human Diseases Detectionmentioning

confidence: 99%

Section: Analysis and Characterization Of Balf Proteomementioning

confidence: 99%

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Human body fluid proteome analysis

2006

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“…Sputum proteins were extracted from a 28-y-old cystic fibrosis patient chronically infected with P. aeruginosa, and were separated by 2D-gel electrophoresis, which had been previously established for the characterization of constituent proteins from complex biological samples, such as protein expression in bronchoalveolar lavage fluid [52,53].…”

Section: Site Selectivity Of Tyrosine Nitration In Proteinsmentioning

confidence: 99%

When is Mass Spectrometry Combined with Affinity Approaches Essential? A Case Study of Tyrosine Nitration in Proteins

Petre

Ulrich

Stumbaum

et al. 2012

J. Am. Soc. Mass Spectrom.

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Tyrosine nitration in proteins occurs under physiologic conditions and is increased at disease conditions associated with oxidative stress, such as inflammation and Alzheimer's disease. Identification and quantification of tyrosine-nitrations are crucial for understanding nitration mechanism(s) and their functional consequences. Mass spectrometry (MS) is best suited to identify nitration sites, but is hampered by low stabilities and modification levels and possible structural changes induced by nitration. In this insight, we discuss methods for identifying and quantifying nitration sites by proteolytic affinity extraction using nitrotyrosine (NT)-specific antibodies, in combination with electrospray-MS. The efficiency of this approach is illustrated by identification of specific nitration sites in two proteins in eosinophil granules from several biological samples, eosinophil-cationic protein (ECP) and eosinophil-derived neurotoxin (EDN). Affinity extraction combined with Edman sequencing enabled the quantification of nitration levels, which were found to be 8 % and 15 % for ECP and EDN, respectively. Structure modeling utilizing available crystal structures and affinity studies using synthetic NT-peptides suggest a tyrosine nitration sequence motif comprising positively charged residues in the vicinity of the NT-residue, located at specific surface-accessible sites of the protein structure. Affinities of Tyr-nitrated peptides from ECP and EDN to NT-antibodies, determined by online bioaffinity-MS, provided nanomolar K D values. In contrast, false-positive identifications of nitrations were obtained in proteins from cystic fibrosis patients upon using NT-specific antibodies, and were shown to be hydroxy-tyrosine modifications. These results demonstrate affinity-mass spectrometry approaches to be essential for unequivocal identification of biological tyrosine nitrations.

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“…One of these techniques, twodimensional difference gel electrophoresis (2D-DIGE) (1,18,19), coupled with matrix assisted laser desorption ionizationtime-of-flight/time-of-flight (MALDI-ToF/ToF) tandem mass spectrometry for protein identification has proved very useful in doing quantitative comparisons of protein expression. 2D-DIGE has enabled the development of other posttranslational modification-specific proteomic methods that allow proteomewide screens of protein modifications.Several investigators have conducted preliminary proteomic surveys of human bronchoalveolar lavage (BAL) fluid in normal individuals (62) as well as in patients with several lung diseases (3,7,49,49,63). Several studies also have examined the carbonylation of BAL proteins in various diseases (47,53,54).…”

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confidence: 99%

Age-related changes in the expression and oxidation of bronchoalveolar lavage proteins in the rat

Umstead¹,

Freeman²,

Chinchilli³

et al. 2009

American Journal of Physiology-Lung Cellular and Molecular Phys

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Umstead TM, Freeman WM, Chinchilli VM, Phelps DS. Age-related changes in the expression and oxidation of bronchoalveolar lavage proteins in the rat. Am J Physiol Lung Cell Mol Physiol 296: L14-L29, 2009. First published October 17, 2008 doi:10.1152/ajplung.90366.2008The incidence and severity of many lung diseases change with age. Some diseases, such as pneumonia, occur with increased frequency in children and the elderly. Proteins obtained by bronchoalveolar lavage (BAL) serve as the first line of defense against inhaled toxins and pathogens. Age-related changes in BAL protein expression and oxidative modification were examined in juvenile (1 mo), young adult (2 mo), and aged (18 mo) F344 rats using two-dimensional difference gel electrophoresis (2D-DIGE), matrix-assisted laser desorption ionization-time of flight/time of flight (MALDI-ToF/ToF) tandem mass spectrometry, and carbonyl immunoblotting. Using 2D-DIGE, we detected 563 protein spots, and MALDI-ToF/ToF identified 204 spots comprising 31 proteins; 21 changed significantly (17 increases) between juvenile and young adult or aged rats, but for 12 of these proteins, levels had a biphasic pattern, and levels in aged rats were less than in young adults. Relative carbonylation was determined by comparison of immunostaining with total protein staining on each oxidized protein blot. We found that aged rats had significantly increased oxidation in 13 proteins compared with juvenile rats. Many of the proteins altered in expression or oxidation level had functions in host defense, redox regulation, and protein metabolism. We speculate that low levels of expression of host defense proteins in juvenile rats and decreases in levels of these proteins between young adult and aged rats may predispose these groups to pneumonia. In addition, we have shown age-related increases in protein oxidation that may compromise host defense function in aged rats.aging; proteomics; host defense; carbonylation; proteolysis AGE IS A MAJOR FACTOR in the incidence of various lung diseases, particularly in the young and the old. Lung diseases, including chronic conditions such as emphysema and pulmonary fibrosis and acute lung diseases such as pneumonia, currently affect more than 35 million people in the U.S. Many of these diseases occur with increased frequency and severity in the elderly. However, pneumonia is also a serious problem in young children, both in developing countries, where it can be the leading cause of childhood mortality (48), and in developed countries, where morbidity is significant but mortality is low (16,43).Age also plays a role in how both humans and laboratory animals respond to pollution and various other insults. Aged organisms have an increased inflammatory response to oxidants, pollutants, and other insults (14,15,27,36,40) and are far more likely to become infected. Indeed, in the U.S., influenza and pneumonia are the fifth leading cause of death in patients over the age of 65 (37). Although it remains unclear why the elderly are more susceptible to pneumon...

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High resolution mass spectrometric alveolar proteomics: Identification of surfactant protein SP‐A and SP‐D modifications in proteinosis and cystic fibrosis patients

Cited by 41 publications

References 36 publications

Human body fluid proteome analysis

Human body fluid proteome analysis

When is Mass Spectrometry Combined with Affinity Approaches Essential? A Case Study of Tyrosine Nitration in Proteins

Age-related changes in the expression and oxidation of bronchoalveolar lavage proteins in the rat

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