Analysis of major intracellular proteins of Aspergillus fumigatus by MALDI mass spectrometry:: Identification and characterisation of an elongation factor 1B protein with glutathione transferase activity
Analysis of major intracellular proteins of Aspergillus fumigatus by MALDI mass spectrometry:: Identification and characterisation of an elongation factor 1B protein with glutathione transferase activity
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DOI：
10.1016/j.bbrc.2006.01.078
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发表时间：
2006-03-24
影响因子：
3.1
通讯作者：
Doyle, S
中科院分区：
文献类型：
--
作者：
Carberry, S;Neville, CM;Doyle, S
烟曲霉是一种公认的人类病原体，特别是在免疫功能低下的个体中。带注释的烟曲霉基因组序列的可用性将显着加速我们对这种生物体的了解。然而，有关烟曲霉蛋白质组的信息有限。在这里，我们采用直接蛋白质组方法（2D-PAGE 和 MALDI-MS）和涉及初始谷胱甘肽亲和层析的亚蛋白质组策略来鉴定烟曲霉中主要参与能量代谢和蛋白质生物合成的 54 种蛋白质。此外，两种新型真核延伸因子蛋白（eEF1B gamma），称为 E1fA 和 B 已被鉴定，并在系统发育上证实属于 GST 样蛋白的 eEF1B gamma 类。其中一种蛋白质 (E1fA) 已被纯化至均质，被鉴定为单体酶（分子量 = 20 kDa；pI = 5.9 和 6.5），并发现使用 CDNB 和 ethacrynic 表现出谷胱甘肽转移酶活性比活性（平均标准差，n = 3）为 3.13 +/- 0.27 和 3.43 +/- 1.0 mu mol/min/mg酸，分别。总体而言，这些数据强调了剖析烟曲霉蛋白质组和阐明烟曲霉新功能的新方法的重要性。 (c) 2006 Elsevier Inc. 保留所有权利。
Aspergillus fumigatus is a recognised human pathogen, especially in immunocompromised individuals. The availability of the annotated A. fumigatus genome sequence will significantly accelerate our understanding of this organism. However, limited information is available with respect to the A. fumigatus proteome. Here, both a direct proteomic approach (2D-PAGE and MALDI-MS) and a sub-proteomic strategy involving initial glutathione affinity chromatography have been deployed to identify 54 proteins from A. fumigatus primarily involved in energy metabolism and protein biosynthesis. Furthermore, two novel eukaryotic elongation factor proteins (eEF1B gamma), termed E1fA and B have been identified and phylogenetically confirmed to belong to the eEF1B gamma class of GST-like proteins. One of these proteins (E1fA) has been purified to homogeneity, identified as a monomeric enzyme (molecular mass = 20 kDa; pI = 5.9 and 6.5), and found to exhibit glutathione transferase activity specific activities (mean standard deviation, n = 3) of 3.13 +/- 0.27 and 3.43 +/- 1.0 mu mol/min/mg, using CDNB and ethacrynic acid, respectively. Overall, these data highlight the importance of new approaches to dissect the proteome of, and elucidate novel functions within, A. fumigatus. (c) 2006 Elsevier Inc. All rights reserved.