PROTEOMIC PROFILING OF MATURE WHITE ADIPOCYTES IDENTIFIES PROTEIN CANDIDATES POTENTIALLY INVOLVED IN INSULIN-STIMULATED GLUCOSE UPTAKE

Abstract

Dysregulation of blood glucose homeostasis can lead to hyperglycemia and type 2 diabetes. Adipose tissue, due to its sensitivity to insulin, initiates Glucose Transporter 4 (GLUT4) translocation to aid in glucose uptake into the cells. Quantitative proteomics was used to profile proteins in primary stromovascular cells compared to primary white adipocytes. Proteomic screening identified 41 proteins that were increased in adipocytes and had a potential link to insulinstimulated glucose uptake. CAP2 and NLRC3 were the two proteins chosen to test whether knockdown of their protein would cause a decrease in overall insulinstimulated glucose uptake. Using siRNA each protein was knocked down and then qPCR was used to confirm the knockdown. From there glucose uptake assays were completed to show changes in non-targeting siRNA cells compared to the siRNA treated cells. The experiments showed significant knockdown of each protein but no changes in glucose uptake. Further experiments are being conducted to continue testing possible candidates involved in insulin-stimulated glucose uptake.