Shotgun proteomic analysis of a chromatophore-enriched preparation from the purple phototrophic bacterium Rhodopseudomonas palustris
Shotgun proteomic analysis of a chromatophore-enriched preparation from the purple phototrophic bacterium Rhodopseudomonas palustris
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DOI：
10.1023/b:pres.0000006752.81486.74
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发表时间：
2003-01-01
影响因子：
3.7
通讯作者：
Beatty, JT
中科院分区：
文献类型：
--
作者：
Fejes, AP;Yi, EC;Beatty, JT
评估了蛋白质组学方法，用于分析光合作用相关蛋白质，这些蛋白质是色素细胞的特征，是源自紫色光养细菌胞质膜的颗粒。将来自沼泽红假单胞菌的纯化色素细胞的蛋白质溶解并用胰蛋白酶消化，以产生通过液相色谱进行分级的肽集合。通过分析肽的串联质谱，并与源自最近确定的沼泽红藻基因组序列的文库进行比较，确定肽序列并将其分配给特定蛋白质。总共检测到300个概率值大于或等于0.9的蛋白质，当最小概率值降低到0.5时，检测到的蛋白质数量增加到345个。反应中心的膜整合蛋白、细胞色素 b/c(1)、光捕获和 ATP 酶复合物用作对照，以评估该方法对疏水蛋白的效果。鉴定出新基因，并暂时指定为编码光合作用相关蛋白。我们的结论是，这种方法是评估新的光合作用相关蛋白质（和基因）可能存在的强大方法，尽管需要其他方法来评估新发现基因的确切功能。
A proteomics approach was evaluated for analysis of photosyntheis-related proteins that are characteristic of chromatophores, particles derived from purple phototrophic bacterial intracytoplasmic membranes. Proteins of purified chromatophores from Rhodopseudomonas palustris were solubilized and digested with trypsin, to create a collection of peptides that were fractionated by liquid chromatography. Peptide sequences were determined and assigned to specific proteins by analysis of tandem mass spectra of peptides, and comparison to a library derived from the recently determined R. palustris genome sequence. A total of 300 proteins were detected with a probability value greater than or equal to0.9, and the number of proteins detected increased to 345 when the minimum probability value was reduced to 0.5. Membrane-integral proteins of the reaction center, cytochrome b/c(1), light-harvesting and ATPase complexes were used as controls to assess how well this approach performs with hydrophobic proteins. New genes were identified, and tentatively designated as encoding photosynthesis-related proteins. We conclude that this approach is a powerful method to evaluate the possible existence of new photosynthesis-related proteins (and genes), although alternative methods are needed to evaluate the exact functions of newly discovered genes.