Ding Yan-Huai R, Hixson Kim K, Giometti Carol S, Stanley Ann, Esteve-Núñez Abraham, Khare Tripti, Tollaksen Sandra L, Zhu Wenhong, Adkins Joshua N, Lipton Mary S, Smith Richard D, Mester Tünde, Lovley Derek R
Department of Microbiology, University of Massachusetts, Amherst, MA 01002, USA.
Biochim Biophys Acta. 2006 Jul;1764(7):1198-206. doi: 10.1016/j.bbapap.2006.04.017. Epub 2006 May 16.
The proteome of Geobacter sulfurreducens, a model for the Geobacter species that predominate in many Fe(III)-reducing subsurface environments, was characterized with ultra high-pressure liquid chromatography and mass spectrometry using accurate mass and time (AMT) tags as well as with more traditional two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). Cells were grown under six different growth conditions in order to enhance the potential that a wide range of genes would be expressed. The AMT tag approach was able to identify a much greater number of proteins than could be detected with the 2-D PAGE approach. With the AMT approach over 3,000 gene products were identified, representing about 90% of the total predicted gene products in the genome. A high proportion of predicted proteins in most protein role categories were detected; the highest number of proteins was identified in the hypothetical protein role category. Furthermore, 91 c-type cytochromes of 111 predicted genes in the G. sulfurreducens genome were identified. Differences in the abundance of cytochromes and other proteins under different growth conditions provided information for future functional analysis of these proteins. These results demonstrate that a high percentage of the predicted proteins in the G. sulfurreducens genome are produced and that the AMT tag approach provides a rapid method for comparing differential expression of proteins under different growth conditions in this organism.
硫还原地杆菌是在许多还原Fe(III)的地下环境中占主导地位的地杆菌属的一个模型,其蛋白质组通过使用精确质量和时间(AMT)标签的超高压液相色谱和质谱以及更传统的二维聚丙烯酰胺凝胶电泳(2-D PAGE)进行了表征。为了提高广泛基因表达的可能性,细胞在六种不同的生长条件下培养。与2-D PAGE方法相比,AMT标签方法能够鉴定出更多的蛋白质。通过AMT方法鉴定出了超过3000种基因产物,约占基因组中预测基因产物总数的90%。在大多数蛋白质功能类别中,预测蛋白质的检测比例很高;在假定蛋白质功能类别中鉴定出的蛋白质数量最多。此外,在硫还原地杆菌基因组中预测的111个基因中的91个c型细胞色素被鉴定出来。不同生长条件下细胞色素和其他蛋白质丰度的差异为这些蛋白质未来的功能分析提供了信息。这些结果表明,硫还原地杆菌基因组中预测的蛋白质有很高比例被表达,并且AMT标签方法为比较该生物体在不同生长条件下蛋白质的差异表达提供了一种快速方法。