Sparre T, Christensen U Bjerre, Mose Larsen P, Fey S J, Wrzesinski K, Roepstorff P, Mandrup-Poulsen T, Pociot F, Karlsen A E, Nerup J
Steno Diabetes Center, Gentofte, Denmark.
Diabetologia. 2002 Nov;45(11):1550-61. doi: 10.1007/s00125-002-0953-z. Epub 2002 Sep 18.
AIMS/HYPOTHESIS: Type I (insulin-dependent) diabetes mellitus is characterized by selective destruction of the insulin producing beta cells. Interleukin-1beta (IL-1beta) modulates the beta-cell function, protein synthesis, energy production and causes apoptosis. We have previously shown changes in the expression of 82 out of 1 815 protein spots detected by two dimensional gel electrophoresis in IL-1beta exposed diabetes prone Bio Breeding (BB-DP) rat islets of Langerhans in vitro. The aim of this study was to identify the proteins in these 82 spots by mass spectrometry and compare these changes with those seen in IL-1beta exposed Wistar Furth (WF) rat islets.
The 82 protein spots, that changed expression after IL-1beta exposure, were all re-identified on preparative gels of 200 000 neonatal WF rat islets, cut out and subjected to mass spectrometry for identification.
Forty-five different proteins were identified from 51 spots and grouped according to function: (i) energy transduction and redox potentials; (ii) glycolytic and Krebs cycle enzymes; (iii) protein, DNA and RNA synthesis, chaperoning and protein folding; (iv) signal transduction, regulation, differentiation and apoptosis; (v) cellular defence; and (vi) other functions. Comparison of IL-1beta exposed BB-DP and WF islets showed common changes in 14 proteins and several proteins influencing similar pathways, suggesting that similar routes in the two strains lead to beta-cell destruction.
CONCLUSION/INTERPRETATION: We demonstrate that proteome analysis is a powerful tool to identify proteins and pathways in BB-DP rat islets exposed to IL-1beta.
目的/假设：I型（胰岛素依赖型）糖尿病的特征是产生胰岛素的β细胞被选择性破坏。白细胞介素-1β（IL-1β）调节β细胞功能、蛋白质合成、能量产生并导致细胞凋亡。我们之前已经表明，在体外暴露于IL-1β的糖尿病易感生物繁殖（BB-DP）大鼠胰岛中，二维凝胶电泳检测到的1815个蛋白质点中有82个的表达发生了变化。本研究的目的是通过质谱鉴定这82个点中的蛋白质，并将这些变化与暴露于IL-1β的Wistar Furth（WF）大鼠胰岛中的变化进行比较。
在200000个新生WF大鼠胰岛的制备凝胶上重新鉴定了82个在暴露于IL-1β后表达发生变化的蛋白质点，将其切下并进行质谱鉴定。
从51个点中鉴定出45种不同的蛋白质，并根据功能进行分组：（i）能量转导和氧化还原电位；（ii）糖酵解和三羧酸循环酶；（iii）蛋白质、DNA和RNA合成、伴侣蛋白和蛋白质折叠；（iv）信号转导、调节、分化和细胞凋亡；（v）细胞防御；以及（vi）其他功能。比较暴露于IL-1β的BB-DP和WF胰岛发现，14种蛋白质有共同变化，还有几种蛋白质影响相似的途径，这表明两个品系中相似的途径导致β细胞破坏。
结论/解读：我们证明蛋白质组分析是鉴定暴露于IL-1β的BB-DP大鼠胰岛中蛋白质和途径的有力工具。
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