Network of Surface-Displayed Glycolytic Enzymes in Mycoplasma pneumoniae and Their Interactions with Human Plasminogen
Network of Surface-Displayed Glycolytic Enzymes in Mycoplasma pneumoniae and Their Interactions with Human Plasminogen
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DOI：
10.1128/iai.01071-15
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发表时间：
2016-03-01
影响因子：
3.1
通讯作者：
Dumke, Roger
中科院分区：
文献类型：
--
作者：
Gruendel, Anne;Pfeiffer, Melanie;Dumke, Roger
在不同的细菌中，主要是胞质和代谢蛋白质的特征在于表面定位和与不同的宿主因子相互作用。这些兼职蛋白质包括糖酵解酶，并且已经假设它们影响致病物种的毒力。研究了在基因组减少和无细胞壁的微生物肺炎支原体（Mycoplasma pneumoniae）中表面展示的糖酵解酶的存在及其与作为重要宿主因子的人纤溶酶原的相互作用。在成功表达了19种糖酵解酶并制备了多克隆抗血清后，利用总蛋白分级、菌落印迹、温和蛋白水解和支原体免疫荧光等方法对支原体细胞中的蛋白质进行了定位。肺炎细胞。8种糖酵解酶，丙酮酸脱氢酶A至C（PdhA-C）、甘油醛-3-磷酸脱氢酶（GapA）、乳酸脱氢酶（Ldh）、磷酸甘油酸脱氢酶（Pgm）、丙酮酸激酶（Pyk）和转酮醇酶（Tkt）被证实为表面表达的，并且都能够与纤溶酶原相互作用。在尿激酶纤溶酶原激活剂和纤溶酶特异性底物D-缬氨酰-亮氨酰-赖氨酸-对硝基苯胺二盐酸盐存在下，与重组蛋白PdhB、GapA和Pyk结合的纤溶酶原转化为纤溶酶。此外，纤溶酶原与重组蛋白PdhB或Pgm的复合物可降解人纤维蛋白原。此外，表面展示的蛋白质（除PdhC）结合到人肺上皮细胞，并通过与抗纤溶酶原细胞的预孵育的相互作用显着减少。我们的研究结果表明，纤溶酶原结合和激活不同的表面定位的糖酵解酶的M。肺炎克雷伯氏菌可能在人类呼吸道的成功和长期定殖中起作用。
In different bacteria, primarily cytosolic and metabolic proteins are characterized as surface localized and interacting with different host factors. These moonlighting proteins include glycolytic enzymes, and it has been hypothesized that they influence the virulence of pathogenic species. The presence of surface-displayed glycolytic enzymes and their interaction with human plasminogen as an important host factor were investigated in the genome-reduced and cell wall-less microorganism Mycoplasma pneumoniae, a common agent of respiratory tract infections of humans. After successful expression of 19 glycolytic enzymes and production of polyclonal antisera, the localization of proteins in the mycoplasma cell was characterized using fractionation of total proteins, colony blot, mild proteolysis and immunofluorescence of M. pneumoniae cells. Eight glycolytic enzymes, pyruvate dehydrogenases A to C (PdhA-C), glyceraldehyde-3-phosphate dehydrogenase (GapA), lactate dehydrogenase (Ldh), phosphoglycerate mutase (Pgm), pyruvate kinase (Pyk), and transketolase (Tkt), were confirmed as surface expressed and all are able to interact with plasminogen. Plasminogen bound to recombinant proteins PdhB, GapA, and Pyk was converted to plasmin in the presence of urokinase plasminogen activator and plasmin-specific substrate D-valyl-leucyl-lysine-p-nitroanilide dihydrochloride. Furthermore, human fibrinogen was degraded by the complex of plasminogen and recombinant protein PdhB or Pgm. In addition, surface-displayed proteins (except PdhC) bind to human lung epithelial cells, and the interaction was reduced significantly by preincubation of cells with antiplasminogen. Our results suggest that plasminogen binding and activation by different surface-localized glycolytic enzymes of M. pneumoniae may play a role in successful and long-term colonization of the human respiratory tract.