Proteomic changes associated with diabetes in the BB-DP rat

Abstract

These studies were structured with the aim of utilizing emerging technologies in two-dimensional (2D) gel electrophoresis and mass spectrometry to evaluate protein expression changes associated with type 1 diabetes. We reasoned that a broad examination of diabetic tissues at the protein level might open up novel avenues of investigation of the metabolic and signaling pathways that are adversely affected in type 1 diabetes. This study compared the protein expression of the liver, heart, and skeletal muscle of diabetes-prone rats and matched control rats by semiquantitative liquid chromatography-mass spectrometry and differential in-gel 2D gel electrophoresis. Differential expression of 341 proteins in liver, 43 in heart, and 9 (2D gel only) in skeletal muscle was detected. These data were assembled into the relevant metabolic pathways affected primarily in liver. Multiple covalent modifications were also apparent in 2D gel analysis. Several new hypotheses were generated by these data, including mechanisms of net cytosolic protein oxidation, formaldehyde generation by the methionine cycle, and inhibition of carbon substrate oxidation via reduction in citrate synthase and short-chain acyl-CoA dehydrogenase. metabolism; acyl carnitine; citrate synthase; methionine; liver; heart; skeletal muscle; mass spectroscopy; two-dimensional gel electrophoresis ALTHOUGH EXTENSIVELY STUDIED, type 1 diabetes remains a major health concern, and much of the pathogenesis still eludes understanding. In the Diabetes Control and Complications Trial, the most strongly regulated subjects still developed complications despite aggressive intervention (35). The fundamental pathology of type 1 diabetes is the loss of pancreatic ␤-cell function and the related release of insulin in the regulation of overall metabolic functions of the body. The complications of type 1 diabetes include dramatically altered energy metabolism and glucose handling, protein oxidation, vascular disease, heart disease, retinopathy, neuropathies, nephropathy, and susceptibility to infections. These results showed that the pathology of diabetes exceeds the simple maintenance of plasma glucose and insulin levels. A more global assessment of the diabetic state may provide useful insight. These studies sought to perform a comprehensive screen of the proteomic changes associated with the type 1 diabetic state, utilizing emerging technology in high-throughput semiquantitative proteomics and two-dimensional (2D) gel electrophoresis to detect protein expression differences as well as possible covalent modifications inherent to the diabetic state.Previous studies have revealed that many enzymes and proteins are under transitional control by insulin (45) that would predictably alter the content of these proteins with the reduction of insulin in type 1 diabetes. The major transcription factors involved in insulin action are adipocyte differentiation and determination factor (ADD)-1/sterol regulatory element binding protein (SREBP)-1/peroxisome proliferator-activated receptor (PP...

Protein composition and function of red and white skeletal muscle mitochondria

“…Proteins listed in Fig. 1, A and D, are those that have been identified many times by our laboratory (4,5,8,20,27,41,45) as well as by Kim et al (30).…”

2D-DIGE to identify proteins associated with gestational diabetes in omental adipose tissue

“…Removal of ROS by antioxidant enzymes plays an important part in limiting this damage, and indeed, antioxidants can prevent diabetic embryopathy in rat models of diabetes in pregnancy (Higa et al 2012). PRX6 is a cytosolic peroxidase (Fisher 2011) that has been demonstrated to protect various tissues, such as skin, lung and cardiac muscle, against acute oxidative insults, and that has been shown to be down-regulated in diabetes (Johnson et al 2009). HSPs are molecular chaperones that are classified into families according to their molecular weight (Ellis 1999, Hwang et al 2009.…”

Diversity of human skeletal muscle in health and disease: Contribution of proteomics

“…Johnson et al investigated by 2DE the protein expression of liver, heart and skeletal muscle of diabetes-prone rats [144]. The study provided interesting observations e.g., the limited involvement of skeletal muscle in this disease.…”