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Mass Spectrometry-Based (GeLC-MS/MS) Comparative Proteomic Analysis of Endoscopically (ePFT) Collected Pancreatic and Gastroduodenal Fluids.

Paulo JA, Kadiyala V, Banks PA, Steen H, Conwell DL - Clin Transl Gastroenterol (2012)

Bottom Line: The secretin-stimulated endoscopic pancreatic function test (ePFT) allows for the safe collection of gastroduodenal and pancreatic fluid from the duodenum.The proteases pepsinogens and gastrin were enriched in gastroduodenal fluid, while common pancreatic enzymes (e.g., aminopeptidase N, chymotrypsin C, elastase-3A, trypsin, and carboxypeptidase A1, and elastase 2B) were found in greater abundance in pancreatic fluid.Similarly for protease inhibitors, members of the cystatin family were exclusive to gastroduodenal fluid, while serpins A11, B4, and D1 were exclusive to pancreatic fluid.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Pathology, Children's Hospital Boston and Harvard Medical School, Boston, Massachusetts, USA [2] Proteomics Center at Children's Hospital Boston, Boston, Massachusetts, USA [3] Center for Pancreatic Disease, Division of Gastroenterology, Hepatology and Endoscopy, Brigham and Women's Hospital and Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.

ABSTRACT

Objectives: The secretin-stimulated endoscopic pancreatic function test (ePFT) allows for the safe collection of gastroduodenal and pancreatic fluid from the duodenum. We test the hypothesis that these endoscopically collected fluids have different proteomes. As such, we aim to show that the ePFT method can be used to collect fluid enriched in pancreatic proteins to test for pancreatic function.

Methods: Gastroduodenal and pancreatic fluid were collected sequentially from chronic pancreatitis patients undergoing an ePFT. Proteins from each fluid type were extracted using previously published optimized methods and subjected to GeLC-MS/MS analysis for protein identification and bioinformatics analysis.

Results: Mass spectrometry analysis identified proteins that were exclusive in either gastroduodenal (46) or pancreatic fluid (234). Subsequent quantitative analysis revealed proteins that were differentially abundant with statistical significance. As expected, proteolytic enzymes and protease inhibitors were among the differentially detected proteins. The proteases pepsinogens and gastrin were enriched in gastroduodenal fluid, while common pancreatic enzymes (e.g., aminopeptidase N, chymotrypsin C, elastase-3A, trypsin, and carboxypeptidase A1, and elastase 2B) were found in greater abundance in pancreatic fluid. Similarly for protease inhibitors, members of the cystatin family were exclusive to gastroduodenal fluid, while serpins A11, B4, and D1 were exclusive to pancreatic fluid.

Conclusions: We have shown that ePFT collection coupled with mass spectrometry can be used to identify differentially detected proteins in gastroduodenal and pancreatic fluids. The data obtained using GeLC-MS/MS techniques provide further evidence supporting the feasibility of using ePFT-collected fluid to study specific diseases of the upper gastrointestinal tract, such as chronic pancreatitis.

No MeSH data available.


Related in: MedlinePlus

Comparing proteins identified by GeLC-MS/MS in gastroduodenal and pancreatic fluid. Qualitative analysis identified proteins exclusive to gastroduodenal and pancreatic fluid. GDF, gastroduodenal fluid; PF, pancreatic fluid.
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fig3: Comparing proteins identified by GeLC-MS/MS in gastroduodenal and pancreatic fluid. Qualitative analysis identified proteins exclusive to gastroduodenal and pancreatic fluid. GDF, gastroduodenal fluid; PF, pancreatic fluid.

Mentions: Using our mass spectrometry-based strategy in which a non-redundant list of proteins were determined using the Scaffold3 software, we identified a total of 285 proteins in gastroduodenal fluid and 473 proteins in pancreatic fluid (Figure 3). Of these proteins, 46 (Supplementary Table 1) were determined to be exclusive to gastroduodenal fluid and 234 (Supplementary Table 2) were determined to be exclusive to pancreatic fluid. Proteins common to both the gastroduodenal and pancreatic fluid samples (239) were analyzed further by QSPEC to determine proteins that were not exclusive to either fluid, but were of enriched in one fluid or the other. In Table 2 we summarized the protein identification data. In addition, Supplementary Figure 1 illustrates the differences in the number of proteins identified in pancreatic and gastroduodenal fluid for each subject.


Mass Spectrometry-Based (GeLC-MS/MS) Comparative Proteomic Analysis of Endoscopically (ePFT) Collected Pancreatic and Gastroduodenal Fluids.

Paulo JA, Kadiyala V, Banks PA, Steen H, Conwell DL - Clin Transl Gastroenterol (2012)

Comparing proteins identified by GeLC-MS/MS in gastroduodenal and pancreatic fluid. Qualitative analysis identified proteins exclusive to gastroduodenal and pancreatic fluid. GDF, gastroduodenal fluid; PF, pancreatic fluid.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3367612&req=5

fig3: Comparing proteins identified by GeLC-MS/MS in gastroduodenal and pancreatic fluid. Qualitative analysis identified proteins exclusive to gastroduodenal and pancreatic fluid. GDF, gastroduodenal fluid; PF, pancreatic fluid.
Mentions: Using our mass spectrometry-based strategy in which a non-redundant list of proteins were determined using the Scaffold3 software, we identified a total of 285 proteins in gastroduodenal fluid and 473 proteins in pancreatic fluid (Figure 3). Of these proteins, 46 (Supplementary Table 1) were determined to be exclusive to gastroduodenal fluid and 234 (Supplementary Table 2) were determined to be exclusive to pancreatic fluid. Proteins common to both the gastroduodenal and pancreatic fluid samples (239) were analyzed further by QSPEC to determine proteins that were not exclusive to either fluid, but were of enriched in one fluid or the other. In Table 2 we summarized the protein identification data. In addition, Supplementary Figure 1 illustrates the differences in the number of proteins identified in pancreatic and gastroduodenal fluid for each subject.

Bottom Line: The secretin-stimulated endoscopic pancreatic function test (ePFT) allows for the safe collection of gastroduodenal and pancreatic fluid from the duodenum.The proteases pepsinogens and gastrin were enriched in gastroduodenal fluid, while common pancreatic enzymes (e.g., aminopeptidase N, chymotrypsin C, elastase-3A, trypsin, and carboxypeptidase A1, and elastase 2B) were found in greater abundance in pancreatic fluid.Similarly for protease inhibitors, members of the cystatin family were exclusive to gastroduodenal fluid, while serpins A11, B4, and D1 were exclusive to pancreatic fluid.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Pathology, Children's Hospital Boston and Harvard Medical School, Boston, Massachusetts, USA [2] Proteomics Center at Children's Hospital Boston, Boston, Massachusetts, USA [3] Center for Pancreatic Disease, Division of Gastroenterology, Hepatology and Endoscopy, Brigham and Women's Hospital and Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.

ABSTRACT

Objectives: The secretin-stimulated endoscopic pancreatic function test (ePFT) allows for the safe collection of gastroduodenal and pancreatic fluid from the duodenum. We test the hypothesis that these endoscopically collected fluids have different proteomes. As such, we aim to show that the ePFT method can be used to collect fluid enriched in pancreatic proteins to test for pancreatic function.

Methods: Gastroduodenal and pancreatic fluid were collected sequentially from chronic pancreatitis patients undergoing an ePFT. Proteins from each fluid type were extracted using previously published optimized methods and subjected to GeLC-MS/MS analysis for protein identification and bioinformatics analysis.

Results: Mass spectrometry analysis identified proteins that were exclusive in either gastroduodenal (46) or pancreatic fluid (234). Subsequent quantitative analysis revealed proteins that were differentially abundant with statistical significance. As expected, proteolytic enzymes and protease inhibitors were among the differentially detected proteins. The proteases pepsinogens and gastrin were enriched in gastroduodenal fluid, while common pancreatic enzymes (e.g., aminopeptidase N, chymotrypsin C, elastase-3A, trypsin, and carboxypeptidase A1, and elastase 2B) were found in greater abundance in pancreatic fluid. Similarly for protease inhibitors, members of the cystatin family were exclusive to gastroduodenal fluid, while serpins A11, B4, and D1 were exclusive to pancreatic fluid.

Conclusions: We have shown that ePFT collection coupled with mass spectrometry can be used to identify differentially detected proteins in gastroduodenal and pancreatic fluids. The data obtained using GeLC-MS/MS techniques provide further evidence supporting the feasibility of using ePFT-collected fluid to study specific diseases of the upper gastrointestinal tract, such as chronic pancreatitis.

No MeSH data available.


Related in: MedlinePlus