Accéder au contenu
Merck

Proteomic Identification of Target Proteins of Thiodigalactoside in White Adipose Tissue from Diet-Induced Obese Rats.

International journal of molecular sciences (2015-06-30)
Hilal Ahmad Parray, Jong Won Yun
RÉSUMÉ

Previously, galectin-1 (GAL1) was found to be up-regulated in obesity-prone subjects, suggesting that use of a GAL1 inhibitor could be a novel therapeutic approach for treatment of obesity. We evaluated thiodigalactoside (TDG) as a potent inhibitor of GAL1 and identified target proteins of TDG by performing comparative proteome analysis of white adipose tissue (WAT) from control and TDG-treated rats fed a high fat diet (HFD) using two dimensional gel electrophoresis (2-DE) combined with MALDI-TOF-MS. Thirty-two spots from a total of 356 matched spots showed differential expression between control and TDG-treated rats, as identified by peptide mass fingerprinting. These proteins were categorized into groups such as carbohydrate metabolism, tricarboxylic acid (TCA) cycle, signal transduction, cytoskeletal, and mitochondrial proteins based on functional analysis using Protein Annotation Through Evolutionary Relationship (PANTHER) and Database for Annotation, Visualization, Integrated Discovery (DAVID) classification. One of the most striking findings of this study was significant changes in Carbonic anhydrase 3 (CA3), Voltage-dependent anion channel 1 (VDAC1), phosphatidylethanolamine-binding protein 1 (PEBP1), annexin A2 (ANXA2) and lactate dehydrogenase A chain (LDHA) protein levels between WAT from control and TDG-treated groups. In addition, we confirmed increased expression of thermogenic proteins as well as reduced expression of lipogenic proteins in response to TDG treatment. These results suggest that TDG may effectively prevent obesity, and TDG-responsive proteins can be used as novel target proteins for obesity treatment.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Glycérol, for molecular biology, ≥99.0%
Sigma-Aldrich
Sodium Dodecyl Sulfate, BioReagent, suitable for electrophoresis, for molecular biology, ≥98.5% (GC)
Sigma-Aldrich
DL-Dithiothréitol solution, BioUltra, for molecular biology, ~1 M in H2O
Sigma-Aldrich
Acétonitrile, anhydrous, 99.8%
Sigma-Aldrich
Sodium Dodecyl Sulfate, ≥99.0% (GC), dust-free pellets
Sigma-Aldrich
Urée, powder, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
Iodoacétamide, Single use vial of 56 mg
Sigma-Aldrich
Iodoacétamide, BioUltra
Supelco
DL-Dithiothréitol solution, 1 M in H2O
Sigma-Aldrich
Sodium Dodecyl Sulfate solution, BioUltra, for molecular biology, 10% in H2O
Sigma-Aldrich
Glycérol, ≥99.5%
Sigma-Aldrich
Sodium Dodecyl Sulfate, ACS reagent, ≥99.0%
Sigma-Aldrich
Iodoacétamide, ≥99% (NMR), crystalline
Sigma-Aldrich
Urée, ACS reagent, 99.0-100.5%
Sigma-Aldrich
Urea solution, BioUltra, ~8 M in H2O
Sigma-Aldrich
Sodium Dodecyl Sulfate, ReagentPlus®, ≥98.5% (GC)
Supelco
Urea, 8 M (after reconstitution with 16 mL high purity water)
Sigma-Aldrich
Glycérol, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for electrophoresis, ≥99% (GC)