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Two-dimensional gel proteome reference map of human small intestine.

Simula MP, Cannizzaro R, Marin MD, Pavan A, Toffoli G, Canzonieri V, De Re V - Proteome Sci (2009)

Bottom Line: Alterations in the bowel reduce its capability to absorb nutrients, minerals and fat-soluble vitamins.Adult celiac disease patients present an overall risk of cancer that is almost twice than that found in the general population.Spots identifications, reported in a database, will be helpful to identify the variability in protein expression levels, in isoforms expression, or in post-translational modifications associated to pathology or to a therapy.

View Article: PubMed Central - HTML - PubMed

Affiliation: CRO Centro di Riferimento Oncologico, IRCCS National Cancer Institute, Aviano, PN, Italy. mpsimula@cro.it

ABSTRACT

Background: The small intestine is an important human organ that plays a central role in many physiological functions including digestion, absorption, secretion and defense. Duodenal pathologies include, for instance, the ulcer associated to Helicobacter Pylori infection, adenoma and, in genetically predisposed individuals, celiac disease. Alterations in the bowel reduce its capability to absorb nutrients, minerals and fat-soluble vitamins. Anemia and osteopenia or osteoporosis may develop as a consequence of vitamins malabsorption. Adenoma is a benign tumor that has the potential to become cancerous. Adult celiac disease patients present an overall risk of cancer that is almost twice than that found in the general population. These disease processes are not completely known.To date, a two dimensional (2D) reference map of proteins expressed in human duodenal tissue is not yet available: the aim of our study was to characterize the 2D protein map, and to identify proteins of duodenal mucosa of adult individuals without duodenal illness, to create a protein database. This approach, may be useful for comparing similar protein samples in different laboratories and for the molecular characterization of intestinal pathologies without recurring to the use of surgical material.

Results: The enrolled population comprised five selected samples (3 males and 2 females, aged 19 to 42), taken from 20 adult subjects, on their first visit at the gastroenterology unit for a suspected celiac disease, who did not turn to be affected by any duodenal pathology after gastrointestinal and histological evaluations. Proteins extracted from the five duodenal mucosal specimens were singly separated by 2D gel electrophoresis. After image analysis of each 2D gel, 179 protein spots, representing 145 unique proteins, from 218 spots tested, were successfully identified by MALDI-TOF ms analysis. Normalized volumes, for each protein, have been reported for every gel. Proteins have been grouped according to their biological/metabolic functions.

Conclusion: This study represents to date the first detailed and reproducible 2D protein map of human duodenum. Spots identifications, reported in a database, will be helpful to identify the variability in protein expression levels, in isoforms expression, or in post-translational modifications associated to pathology or to a therapy.

No MeSH data available.


Related in: MedlinePlus

Proteins of human duodenum tissue identified by MALDI-TOF MS from the 2-D gel.
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Figure 2: Proteins of human duodenum tissue identified by MALDI-TOF MS from the 2-D gel.

Mentions: The included protein spots obtained from all five 2D gels, ranged from 822 to 1130. All gels were matched to the duodenum master 2D map and the normal volume of each protein spot, and for each sample, was determined by DeCyder software. The Coomassie-stained preparative gel image was then matched with analytical DIGE gel images and the visible spots, typically containing enough protein for mass spectrometry analysis, were selected for identification. A total of 179 protein spots, representing 145 unique proteins, have been so far positively identified and are illustrated in Figures 1, 2, 3, 4; quantitative data are reported on Additional file 2. We failed in identifying 39 spots. As regards faint spots the low protein amount resulted in low sequence coverage that was not sufficient for certain identifications. In the remaining 11 spots we failed in obtaining certain identifications due to the intrinsic limits of PMF: the possibility to identify only proteins for which sequences are already known and the inability to detect post-translational modifications and protein fragments.


Two-dimensional gel proteome reference map of human small intestine.

Simula MP, Cannizzaro R, Marin MD, Pavan A, Toffoli G, Canzonieri V, De Re V - Proteome Sci (2009)

Proteins of human duodenum tissue identified by MALDI-TOF MS from the 2-D gel.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2667413&req=5

Figure 2: Proteins of human duodenum tissue identified by MALDI-TOF MS from the 2-D gel.
Mentions: The included protein spots obtained from all five 2D gels, ranged from 822 to 1130. All gels were matched to the duodenum master 2D map and the normal volume of each protein spot, and for each sample, was determined by DeCyder software. The Coomassie-stained preparative gel image was then matched with analytical DIGE gel images and the visible spots, typically containing enough protein for mass spectrometry analysis, were selected for identification. A total of 179 protein spots, representing 145 unique proteins, have been so far positively identified and are illustrated in Figures 1, 2, 3, 4; quantitative data are reported on Additional file 2. We failed in identifying 39 spots. As regards faint spots the low protein amount resulted in low sequence coverage that was not sufficient for certain identifications. In the remaining 11 spots we failed in obtaining certain identifications due to the intrinsic limits of PMF: the possibility to identify only proteins for which sequences are already known and the inability to detect post-translational modifications and protein fragments.

Bottom Line: Alterations in the bowel reduce its capability to absorb nutrients, minerals and fat-soluble vitamins.Adult celiac disease patients present an overall risk of cancer that is almost twice than that found in the general population.Spots identifications, reported in a database, will be helpful to identify the variability in protein expression levels, in isoforms expression, or in post-translational modifications associated to pathology or to a therapy.

View Article: PubMed Central - HTML - PubMed

Affiliation: CRO Centro di Riferimento Oncologico, IRCCS National Cancer Institute, Aviano, PN, Italy. mpsimula@cro.it

ABSTRACT

Background: The small intestine is an important human organ that plays a central role in many physiological functions including digestion, absorption, secretion and defense. Duodenal pathologies include, for instance, the ulcer associated to Helicobacter Pylori infection, adenoma and, in genetically predisposed individuals, celiac disease. Alterations in the bowel reduce its capability to absorb nutrients, minerals and fat-soluble vitamins. Anemia and osteopenia or osteoporosis may develop as a consequence of vitamins malabsorption. Adenoma is a benign tumor that has the potential to become cancerous. Adult celiac disease patients present an overall risk of cancer that is almost twice than that found in the general population. These disease processes are not completely known.To date, a two dimensional (2D) reference map of proteins expressed in human duodenal tissue is not yet available: the aim of our study was to characterize the 2D protein map, and to identify proteins of duodenal mucosa of adult individuals without duodenal illness, to create a protein database. This approach, may be useful for comparing similar protein samples in different laboratories and for the molecular characterization of intestinal pathologies without recurring to the use of surgical material.

Results: The enrolled population comprised five selected samples (3 males and 2 females, aged 19 to 42), taken from 20 adult subjects, on their first visit at the gastroenterology unit for a suspected celiac disease, who did not turn to be affected by any duodenal pathology after gastrointestinal and histological evaluations. Proteins extracted from the five duodenal mucosal specimens were singly separated by 2D gel electrophoresis. After image analysis of each 2D gel, 179 protein spots, representing 145 unique proteins, from 218 spots tested, were successfully identified by MALDI-TOF ms analysis. Normalized volumes, for each protein, have been reported for every gel. Proteins have been grouped according to their biological/metabolic functions.

Conclusion: This study represents to date the first detailed and reproducible 2D protein map of human duodenum. Spots identifications, reported in a database, will be helpful to identify the variability in protein expression levels, in isoforms expression, or in post-translational modifications associated to pathology or to a therapy.

No MeSH data available.


Related in: MedlinePlus