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Original

Identification of preferential protein targets for carbonylation in human mature adipocytes treated with native or glycated albumin

, , &
Pages 1078-1088 | Received 14 Jan 2007, Published online: 07 Jul 2009
 

Abstract

Oxidative modifications in proteins can participate in the regulation of cellular functions and are frequently observed in numerous states of diseases. Albumin can undergo increased glycation during diabetes. An accumulation of oxidatively modified proteins in human mature adipocytes incubated with glycated albumin has previously been described. This study herein reports the identification of specifically carbonylated targets following separation of the cell proteins by 2D gels, Western blotting and mass spectrometry analyses. It identified eight oxidatively modified proteins, two of which (ACTB and Annexin A2) appeared as significantly more carbonylated in adipocytes treated with glycated albumin than with native albumin. Intracellular stress, evaluated in SW872 cell line, showed an impairment in the protective antioxidant action exerted by native BSA after the glycation of the protein. Decreased proteasome peptidase activities were found in glycated BSA-treated mature adipocytes. The data suggest an association of oxidative damage with the progression of diabetes disorders at the adipocytes level.

Abbreviations
ACTB=

Actin Beta

AGEs=

Advanced Glycated Endproducts

BSA=

Bovine Serum Albumin

BSAGX=

BSA incubated with X mm of glucose

BSAG0=

BSA incubated in the absence of glucose

ECL=

Enhanced Luminol Chemiluminescence

LDH=

Lactate Deshydrogenase

MAD2=

mitochondrial aldehyde dehydrogenase 2, OS

Oxidative Stress=

PAGE, Polyacrylamide Gel Electrophoresis

RAGE=

Receptor for Advanced Glycated Endproducts

ROS=

Reactive Oxygen Species

Abbreviations
ACTB=

Actin Beta

AGEs=

Advanced Glycated Endproducts

BSA=

Bovine Serum Albumin

BSAGX=

BSA incubated with X mm of glucose

BSAG0=

BSA incubated in the absence of glucose

ECL=

Enhanced Luminol Chemiluminescence

LDH=

Lactate Deshydrogenase

MAD2=

mitochondrial aldehyde dehydrogenase 2, OS

Oxidative Stress=

PAGE, Polyacrylamide Gel Electrophoresis

RAGE=

Receptor for Advanced Glycated Endproducts

ROS=

Reactive Oxygen Species

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