Djordjevic Michael A
Australian Research Council Centre of Excellence for Integrative Legume Research, Genomic interactions Group, Research School of Biological Sciences, Australian National University, GPO Box 475, Canberra, ACT 2601, Australia.
Proteomics. 2004 Jul;4(7):1859-72. doi: 10.1002/pmic.200300802.
The proteome of the model symbiotic bacterium, Sinorhizobium meliloti was examined to determine the enzymatic reactions and cell processes that occur when S. meliloti occupies the root nodules of Medicago truncatula and Melilotus alba. The proteomes of the nodule bacteria were compared to that of S. meliloti grown under laboratory cultured conditions as an additional control. All the detectable protein spots on the two-dimensional (2-D) gels between pH 4-7 were analyzed. In total, the identity of proteins in 1545 spots from 2-D gels was determined using peptide mass fingerprinting. There were clear differences in the proteome of nodule bacteria and cultured bacteria and putative nodule-specific and nodule suppressed proteins were identified. The data were analyzed using metabolic pathway prediction programs and used to review the biochemical and genetic studies that had been done previously on S. meliloti over several decades. There was a broad congruency between the proteomic and biochemical data when the overall pathways of central carbon and nitrogen metabolism were considered. A selective suite of ABC-type transporters was present in nodule bacteria that were biased towards the transport of amino acids and inorganic ions (P and Fe) suggesting that a highly specialized nutrient exchange was occurring between the nodule bacteria and the host. Proteins prominent in nodule bacteria were those involved in the pathways for vitamin synthesis and stress-related processes (chaperoning, heat shock, detoxification of reactive oxygen species, regulation of stress and osmo-regulation). Some of these proteins were found only in nodule bacteria. These results show the extent of the shift in metabolism that occurs when S. meliloti invades legume plants and establishes a nitrogen fixing symbiosis.
对模式共生细菌苜蓿中华根瘤菌的蛋白质组进行了检测,以确定苜蓿中华根瘤菌占据截形苜蓿和白花草木樨根瘤时发生的酶促反应和细胞过程。将根瘤细菌的蛋白质组与在实验室培养条件下生长的苜蓿中华根瘤菌的蛋白质组进行比较,作为额外的对照。分析了pH 4-7范围内二维(2-D)凝胶上所有可检测到的蛋白质斑点。总共使用肽质量指纹图谱确定了二维凝胶上1545个斑点中蛋白质的身份。根瘤细菌和培养细菌的蛋白质组存在明显差异,并鉴定出了假定的根瘤特异性和根瘤抑制蛋白。使用代谢途径预测程序对数据进行分析,并用于回顾几十年来先前对苜蓿中华根瘤菌进行的生化和遗传研究。当考虑中心碳和氮代谢的总体途径时,蛋白质组学数据和生化数据之间存在广泛的一致性。根瘤细菌中存在一组选择性的ABC型转运蛋白,这些转运蛋白偏向于氨基酸和无机离子(磷和铁)的转运,这表明根瘤细菌与宿主之间正在发生高度专业化的营养交换。在根瘤细菌中突出的蛋白质是那些参与维生素合成途径和应激相关过程(伴侣蛋白、热休克、活性氧解毒应激调节和渗透调节)的蛋白质。其中一些蛋白质仅在根瘤细菌中发现。这些结果表明,苜蓿中华根瘤菌侵入豆科植物并建立固氮共生关系时,代谢发生了转变。