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  • A proteomics approach to detect tissue-wide adaptive changes in the pancreas associated with increased pancreatic α-amylase activity in domestic cattle (Bos taurus).

A proteomics approach to detect tissue-wide adaptive changes in the pancreas associated with increased pancreatic α-amylase activity in domestic cattle (Bos taurus).

Comparative biochemistry and physiology. Part D, Genomics & proteomics (2013-01-01)
Simone Holligan, Jiaxi Wang, John P Cant, Kendall C Swanson
ABSTRACT

We used a proteomics-based approach to investigate potential regulatory proteins in the pancreas of domestic cattle (Bos taurus) that were associated with differences in pancreatic α-amylase activity. Two groups of 48 and 45 crossbred steers in years 1 and 2, respectively, were fed a high moisture corn-based diet and were ranked according to their pancreatic α-amylase activity. Steers (n=18) with high, medium, and low α-amylase activity were selected, with 3 for each activity range and 9 for each experimental year, and their proteomic profiles were compared. Pancreatic samples from each animal were fractionated using 2D-HPLC and fractions detected using UV spectrophotometry. Software analysis revealed 119 common protein fractions among the 18 animals, and statistical analysis revealed 10 of these fractions differing (P<0.10) in abundance between animals from the high and low pancreatic α-amylase activity groups. Five protein fractions identified after tandem mass spectrometry analysis and database searches were found to match proteins with protein-binding, nucleotide/DNA-binding or enzymatic capabilities. Bioinformatics analysis of these fractions revealed porphobilinogen deaminase, a DNA-binding protein, and a putative S1 peptidase that increased in abundance with increasing α-amylase activity; with a putative ATP/GTP binding protein decreasing in abundance with increasing pancreatic α-amylase activity. Changes in these fractions may represent adaptations of the pancreas in domestic cattle that are associated with differences in α-amylase activity.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
α-Amylase from Bacillus sp., powder, yellow-brown, ~50 U/mg
Sigma-Aldrich
α-Amylase from Bacillus sp., powder, yellow-brown, ~380 U/mg
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α-Amylase from porcine pancreas, Type VI-B, ≥5 units/mg solid
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Taka-Diastase from Aspergillus oryzae, powder, slightly beige, ~100 U/mg
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α-Amylase from Bacillus sp., Type II-A, lyophilized powder, ≥1,500 units/mg protein (biuret)
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α-Amylase from Bacillus sp., powder, ≥400 units/mg protein (Lowry)
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α-Amylase from porcine pancreas, PMSF Treated, Type I-A, saline suspension, ≥1000 units/mg protein (E1%/280)
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α-Amylase, heat-stable, solution, for use in Total Dietary Fiber Assay, TDF-100A
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α-Amylase from porcine pancreas, Type I-A, PMSF treated, saline suspension, 700-1400 units/mg protein (E1%/280)
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α-Amylase from Bacillus sp., liquid
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α-Amylase from Aspergillus oryzae, powder, ~1.5 U/mg (~0.2 U acc. to Willstätter)
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α-Amylase from Aspergillus oryzae, ≥150 units/mg protein (biuret)
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α-Amylase from Aspergillus oryzae, aqueous solution, ≥800 FAU/g
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α-Amylase from Bacillus licheniformis, Type XII-A, saline solution, ≥500 units/mg protein (biuret)
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α-Amylase from Bacillus licheniformis, lyophilized powder, 500-1,500 units/mg protein, 93-100% (SDS-PAGE)
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α-Amylase from human saliva, Type XIII-A, lyophilized powder, 300-1,500 units/mg protein
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α-Amylase from Aspergillus oryzae, powder, ~30 U/mg
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