Barry Richard C, Young Mark J, Stedman Kenneth M, Dratz Edward A
Biological Monitoring and Modeling, Pacific NW National Laboratory, Richland, WA, USA.
Electrophoresis. 2006 Jul;27(14):2970-83. doi: 10.1002/elps.200500851.
A proteomic map of Sulfolobus solfataricus P2, an archaeon that grows optimally at 80 degrees C and pH 3.2, was developed using high-resolution 2-DE and peptide mass fingerprinting. A total of 867 protein spots (659 aqueous Tris-soluble spots and 208 aqueous Tris-insoluble) were mapped over IPG 3-10, 4-7, and 6-11, with second-dimensional gels made of 8-18% polyacrylamide. Three hundred and twenty-four different gene products were represented by the 867 spots, with 274 gene products being identified in the Tris-soluble fractions and 100 gene products in the Tris-insoluble portion. Fifty gene products were found on gels from both fractions. Additionally, an average of 1.50 +/- 0.12 isoforms/protein was identified. This mapping study confirmed the expression of proteins involved in numerous metabolic, transport, energy production, nucleic acid replication, translation, and transcription pathways. Of particular interest, phosphoenolpyruvate carboxykinase (SSO2537) was detected even though the pathway for gluconeogenesis is unknown for this archaeon. Tris-soluble fractions contained many cytosolic proteins while Tris-insoluble fractions contained many membrane-associated proteins, including ABC transporters and an ATP synthase. This study provides an optimized 2-DE approach for investigating the biochemical pathways and post-translational modifications employed by Sulfolobus to survive in its extreme environment.
嗜热栖热菌P2是一种在80摄氏度和pH值3.2的条件下生长最佳的古生菌，利用高分辨率二维电泳和肽质量指纹图谱技术绘制了其蛋白质组图谱。在IPG 3 - 10、4 - 7和6 - 11范围内共定位了867个蛋白质点（659个水溶性Tris可溶点和208个水溶性Tris不溶点），第二维凝胶由8 - 18%的聚丙烯酰胺制成。867个点代表了324种不同的基因产物，其中274种基因产物在Tris可溶部分被鉴定出来，100种基因产物在Tris不溶部分被鉴定出来。在两个部分的凝胶上都发现了50种基因产物。此外，平均每个蛋白质鉴定出1.50±0.12种异构体。这项图谱研究证实了参与众多代谢、运输、能量产生、核酸复制、翻译和转录途径的蛋白质的表达。特别值得关注的是，尽管这种古生菌的糖异生途径尚不清楚，但仍检测到磷酸烯醇式丙酮酸羧激酶（SSO2537）。Tris可溶部分包含许多胞质蛋白，而Tris不溶部分包含许多与膜相关的蛋白，包括ABC转运蛋白和一种ATP合酶。这项研究为研究嗜热栖热菌在其极端环境中生存所采用的生化途径和翻译后修饰提供了一种优化的二维电泳方法。
