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Dynamic regulation of uncoupling protein 2 content in INS-1E insulinoma cells
Authors:Vian Azzu  Charles Affourtit  Eamon P. Breen  Nadeene Parker  Martin D. Brand
Affiliation:MRC Dunn Human Nutrition Unit, Wellcome Trust/MRC Building, Hills Road, Cambridge, CB2 0XY, UK
Abstract:Uncoupling protein 2 (UCP2) regulates glucose-stimulated insulin secretion in pancreatic beta-cells. UCP2 content, measured by calibrated immunoblot in INS-1E insulinoma cells (a pancreatic beta-cell model) grown in RPMI medium, and INS-1E mitochondria, was 2.0 ng/million cells (7.9 ng/mg mitochondrial protein). UCP2 content was lower in cells incubated without glutamine and higher in cells incubated with 20 mM glucose, and varied from 1.0-4.4 ng/million cells (2.7-14.5 ng/mg mitochondrial protein). This dynamic response to nutrients was achieved by varied expression rates against a background of a very short UCP2 protein half-life of about 1 h.
Keywords:ANT, adenine nucleotide translocase   GSIS, glucose-stimulated insulin secretion   FCS, fetal calf serum   Q, glutamine   ROS, reactive oxygen species   UCP, uncoupling protein   rUCP2, recombinant UCP2
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