Department of General Surgery, the First Affiliated Hospital, Nanjing Medical University, Nanjing, People's Republic of China.
PLoS One. 2013;8(1):e54489. doi: 10.1371/journal.pone.0054489. Epub 2013 Jan 17.
Nucleation of cholesterol monohydrate crystals following the aggregation and fusion of cholesterol-enriched vesicles is a critical procedure in the formation of cholesterol gallstone. Biliary proteins play important roles in the process. It is inefficient to screen pro-nucleating or anti-nucleating proteins with routine physiochemical techniques, by which we discovered several pro-nucleating proteins.
METHODOLOGY/PRINCIPAL FINDINGS: Based on comparative proteomic technologies, we investigated the differentially expressed proteins between the cholesterol gallstone and control groups, and between the vesicular phase and micellar phase. There are 401±75 spots detected on the cholesterol gallstone group and 389±94 spots on the control group gels, 120±24 spots detected on vesicular phase and 198±37 on micellar phase gels, and accordingly 22 and 8 differentially expressed proteins were identified successfully, respectively. Three of them, HSA, Profilin and Retinol Binding Protein, were validated by Western blot.
CONCLUSION/SIGNIFICANCE: Some of the identified proteins are in good agreement with proteins reported to be involved in the gallstone formation before. The information from this study might provide some important clues to uncover the key proteins involved in the formation of cholesterol gallstone.
胆固醇一水合物晶体在富含胆固醇的囊泡聚集和融合后成核是胆固醇胆结石形成的关键步骤。胆盐蛋白在该过程中起重要作用。用常规理化技术筛选成核前体或抗成核蛋白的效率不高，通过这种方法我们发现了几种成核前体蛋白。
方法/主要发现：基于比较蛋白质组学技术，我们研究了胆固醇胆结石组与对照组之间以及囊泡相和胶束相之间差异表达的蛋白质。在胆固醇胆结石组凝胶上检测到 401±75 个斑点，在对照组凝胶上检测到 389±94 个斑点，在囊泡相凝胶上检测到 120±24 个斑点，在胶束相凝胶上检测到 198±37 个斑点，分别成功鉴定出 22 和 8 种差异表达蛋白。其中三种蛋白，HSA、原肌球蛋白和视黄醇结合蛋白，通过 Western blot 进行了验证。
结论/意义：鉴定出的一些蛋白与之前报道的参与胆结石形成的蛋白相符。本研究提供的信息可能为揭示胆固醇胆结石形成过程中涉及的关键蛋白提供了一些重要线索。
