Limits...
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

Gene ontology (GO) analysis revealed the molecular function of proteins that were exclusive to or enriched in gastroduodenal or pancreatic fluid. Using Scaffold3, the two sets of differentially detected proteins were used for GO classification of molecular function. GDF, gastroduodenal fluid; PF, pancreatic fluid.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3367612&req=5

fig4: Gene ontology (GO) analysis revealed the molecular function of proteins that were exclusive to or enriched in gastroduodenal or pancreatic fluid. Using Scaffold3, the two sets of differentially detected proteins were used for GO classification of molecular function. GDF, gastroduodenal fluid; PF, pancreatic fluid.

Mentions: Using Scaffold3, we investigated the molecular function of the differentially detected proteins (Figure 4). We noted that a large percentage of the proteins identified from both fluids were classified as binding, catalytic, and enzyme regulators. The binding category is very broad and includes ion, protein, lipid, nucleotide, carbohydrate, and oxygen binding. As such, it is expected that this category would be relatively large. The catalytic and enzyme regulator categories were of particular interest in regard to proteases and protease inhibitors related to digestion. Table 3 lists proteases that were exclusive to a particular fluid, while Table 4 lists those proteases that were enriched, to a statistically significant degree, in a particular fluid. Similarly, for the identified protease inhibitors, Table 5 lists those that were exclusive to a particular fluid, whereas Table 6 lists those that were enriched in a particular fluid.


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)

Gene ontology (GO) analysis revealed the molecular function of proteins that were exclusive to or enriched in gastroduodenal or pancreatic fluid. Using Scaffold3, the two sets of differentially detected proteins were used for GO classification of molecular function. GDF, gastroduodenal fluid; PF, pancreatic fluid.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: Gene ontology (GO) analysis revealed the molecular function of proteins that were exclusive to or enriched in gastroduodenal or pancreatic fluid. Using Scaffold3, the two sets of differentially detected proteins were used for GO classification of molecular function. GDF, gastroduodenal fluid; PF, pancreatic fluid.
Mentions: Using Scaffold3, we investigated the molecular function of the differentially detected proteins (Figure 4). We noted that a large percentage of the proteins identified from both fluids were classified as binding, catalytic, and enzyme regulators. The binding category is very broad and includes ion, protein, lipid, nucleotide, carbohydrate, and oxygen binding. As such, it is expected that this category would be relatively large. The catalytic and enzyme regulator categories were of particular interest in regard to proteases and protease inhibitors related to digestion. Table 3 lists proteases that were exclusive to a particular fluid, while Table 4 lists those proteases that were enriched, to a statistically significant degree, in a particular fluid. Similarly, for the identified protease inhibitors, Table 5 lists those that were exclusive to a particular fluid, whereas Table 6 lists those that were enriched in a particular fluid.

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