Differential Proteomic Shotgun Analysis Elucidates Involvement of Water Channel Aquaporin 8 in Presence of α-Amylase in the Colon
Differential Proteomic Shotgun Analysis Elucidates Involvement of Water Channel Aquaporin 8 in Presence of α-Amylase in the Colon
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DOI：
10.1021/pr100789v
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发表时间：
2010-12-01
影响因子：
4.4
通讯作者：
Yamamoto, Tadashi
中科院分区：
文献类型：
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作者：
Magdeldin, Sameh;Li, Huiping;Yamamoto, Tadashi
水通道蛋白（AQP）家族在液体的分泌和吸收中起着关键作用，特别是在消化系统和分泌腺中。据报道，在该家族中，AQP 8广泛表达于消化道、肝脏和胰腺的上皮细胞中。在凝胶胰蛋白酶消化与宏嵌入式电子表格分析和采用基于LC MS比对的方法进行胰蛋白酶溶液消化，我们比较了野生型和AQP 8敲除小鼠结肠蛋白质组，实验揭示了α-淀粉酶2在AQP 8缺失小鼠模型中的下调在转录和翻译水平上的验证证实了AQP 8参与α-淀粉酶2的调节。考虑到AQP 8作为水和溶质转运蛋白的重要作用，其在调节结肠上皮细胞的α-淀粉酶2合成中也可能是重要的。在此，我们还证明了我们的耦合方法在选择最可靠和最重要的候选者方面的能力，这是一个在复杂标本和组织提取物中初步筛选生物标志物的适用过程
Aquaporin (AQP) family plays a pivotal role in fluid secretion and absorption especially in the digestive system and secretory glands Within this family AQP8 was reported to be widely expressed in the epithelia of the digestive tract, liver, and pancreas In two parallel experimental platforms with different analytical and comparative approaches, in gel tryptic digestion with macro-embedded spreadsheet analysis and in-solution tryptic digestion with LC MS alignment based approach we compared wild type and AQP8 knockout mice colon proteomes Shared result between both experiments revealed down-regulation of alpha-amylase 2 in AQP8-deleted mice model Verification on both transcriptional and translational levels confirmed the involvement of AQP8 in alpha-amylase 2 regulation Given the profound role of AQP8 as a water and solutes transporter, it might be important in modulating a-amylase 2 synthesis by colonic epithelial cells as well Here, we also proved the capability of our coupled approaches for selecting the most reliable and significant candidates, an applicable process for initial screening of biological biomarkers in complex specimens and tissue extracts