Towards a phosphoproteome map of Corynebacterium glutamicum
Towards a phosphoproteome map of Corynebacterium glutamicum
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DOI：
10.1002/pmic.200300494
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发表时间：
2003-08-01
期刊：
影响因子：
3.4
通讯作者：
Hermann, T
中科院分区：
文献类型：
--
作者：
Bendt, AK;Burkovski, A;Hermann, T
本研究采用两种不同的检测方法研究了革兰氏阳性菌棒状杆菌/分枝杆菌/诺卡菌（CMN）组中重要的工业土壤细菌谷氨酰胺棒状杆菌的磷蛋白组：一种是用[P-33]-磷酸进行体内放射性标记，随后进行放射自显影；另一种是用磷氨基酸特异性单克隆抗体进行免疫染色。经二维凝胶电泳（2-DE），可见约60个[P-33]标记的蛋白点，检测到约90个抗体修饰的蛋白点；两种方法均检测到31个蛋白斑点。通过多肽质量指纹图谱鉴定出41种不同的蛋白，分别为5-烯醇丙酮酰石草酸3-磷酸合成酶、乌头酸酶、酰基辅酶a羧化酶、酰基辅酶a合成酶、ATP （α链和β链合成酶）、氨甲酰磷酸合成酶、柠檬酸合成酶、半胱氨酸合成酶、DnaK、延伸因子G、P、Ts和Tu、烯醇化酶、果糖二磷酸醛缩酶、延霉酶、Gap脱氢酶、谷氨酰胺合成酶I、甘氨酸羟甲基转移酶、GroEL2、GTPase、热诱导转录抑制因子DnaJ2、无机焦磷酸酶、异柠檬酸脱氢酶、酮酸还原异构酶、乳酸脱氢酶、亮氨酸- trna连接酶、脂酰胺脱氢酶、蛋氨酸合成酶、o -乙酰高丝氨酸巯基化酶、丙酮酸羧化酶、丙酮酸激酶、丙酮酸氧化酶、核糖体蛋白S1、RNA聚合酶（β -亚基）、琥珀酰辅酶a：辅酶a转移酶、转酮醇酶和udp - n -乙酰muramyl-丙氨酸连接酶、除了一个假设的35k蛋白和一个假设的葡萄糖激酶。这两种检测技术都用于创建磷蛋白质组图。此外，还研究了氮剥夺对谷氨酰胺磷蛋白组的影响。
In this study, the phosphoproteome of Corynebacterium glutamicum, an industrially important soil bacterium of the Corynebacterium/Mycobacterium/Nocardia (CMN) group of Gram-positive bacteria, was investigated by two different detection methods: first, by in vivo radio-labeling using [P-33]-phosphoric acid with subsequent autoradiography and second, by immunostaining with phosphoamino acid-specific monoclonal antibodies. After two-dimensional gel electrophoresis (2-DE), around 60 [P-33]-labeled protein spots were visualized and around 90 antibody-decorated protein spots detected; 31 of the protein spots were detected with both methods. By peptide mass fingerprinting, 41 different proteins were identified, namely 5-enolpyruvylshikimate 3-phosphate synthase, aconitase, acyl-CoA carboxylase, acyl-CoA synthetase, ATP (synthase alpha- and beta-chain), carbamoyl-phosphate synthase, citrate synthase, cysteine synthase, DnaK, the elongation factors G, P, Ts and Tu, enolase, fructose bisphosphate aldolase, fumarase, Gap dehydrogenase, glutamine synthetase I, glycine hydroxymethyltransferase, GroEL2, GTPase, heat-inducible transcriptional repressor DnaJ2, inorganic pyrophosphatase, isocitrate dehydrogenase, ketol-acid reductoisomerase, lactate dehydrogenase, leucine-tRNA ligase, lipoamide dehydrogenase, methionine synthase, O-acetylhomoserine sulfhydrylase, pyruvate carboxylase, pyruvate kinase, pyruvate oxidase, ribosomal protein S1, RNA polymerase (beta-subunit), succinyl-CoA:CoA transferase, transketolase and UDP-N-acetylmuramyl-L-alanine ligase, besides a hypothetical 35k protein and a hypothetical glucose kinase. Both detection techniques were used to create a phosphoproteome map. Additionally, the influence of nitrogen deprivation on the phosphoproteome of C. glutamicum was investigated.