Substrate specificity and function of acetylpolyamine amidohydrolases from Pseudomonas aeruginosa.
Substrate specificity and function of acetylpolyamine amidohydrolases from Pseudomonas aeruginosa.
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DOI：
10.1186/s12858-016-0063-z
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发表时间：
2016-03-09
期刊：
影响因子：
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通讯作者：
Meyer-Almes FJ
中科院分区：
文献类型：
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作者：
Krämer A;Herzer J;Overhage J;Meyer-Almes FJ
铜绿假单胞菌是一种革兰氏阴性需氧球杆菌属细菌，是一种机会性人类病原体，也是全世界医院获得性感染的第四大常见原因，这些感染通常具有高死亡率，例如呼吸机相关肺炎。铜绿假单胞菌的多胺代谢，特别是乙酰多胺的脱乙酰化反应，至今研究甚少。其他细菌病原体的结果，例如，Y.鼠疫表明多胺可能参与生物膜的形成或赋予对某些抗生素的抗性。为了更详细地阐明乙酰多胺及其酶促脱乙酰化的作用，已经以酶促活性形式表达了来自铜绿假单胞菌的所有三种推定的乙酰多胺酰胺水解酶（APAH）。APAH PA 0321和PA 1409被证明是真正的多胺脱乙酰酶，而PA 3774不能使乙酰化的多胺脱乙酰化。每个APAH都可以水解三氟乙酰化赖氨酸衍生物，但只有PA 1409和更有效的PA 3774也可以处理普通乙酰化赖氨酸底物。铜绿假单胞菌在葡萄糖饥饿下能够利用乙酰尸胺和乙酰腐胺作为碳源。如果PA 0321或PA 1409基因被破坏，而不是PA 3774基因被破坏，铜绿假单胞菌的生长就会减少和延迟。此外，我们能够表明APAH抑制剂SAHA和SATFMK在PA 14和PAO 1野生型菌株中诱导生物膜形成。铜绿假单胞菌具有两种功能性APAH，PA 0321和PA 1409，其使得能够利用乙酰多胺用于铜绿假单胞菌的代谢。相反，预测的APAH，PA 3774的生理作用仍有待阐明。其对合成乙酰化赖氨酸底物进行脱乙酰化的能力表明了蛋白质对未知底物的脱乙酰化功能。本文的在线版本（doi：10.1186/s12858-016-0063-z）包含补充材料，可供授权用户使用。
Pseudomonas aeruginosa, a Gram-negative, aerobic coccobacillus bacterium is an opportunistic human pathogen and worldwide the fourth most common cause of hospital-acquired infections which are often high mortality such as ventilator-associated pneumoniae. The polyamine metabolism of P. aeruginosa and particularly the deacetylation of acetylpolyamines has been little studied up to now. Results with other bacterial pathogens e.g., Y. pestis suggest that polyamines may be involved in the formation of biofilms or confer resistance against certain antibiotics. To elucidate the role of acetylpolyamines and their enzymatic deacetylation in more detail, all three putative acetylpolyamine amidohydrolases (APAHs) from P. aeruginosa have been expressed in enzymatic active form. The APAHs PA0321 and PA1409 are shown to be true polyamine deacetylases, whereas PA3774 is not able to deacetylate acetylated polyamines. Every APAH can hydrolyze trifluoroacetylated lysine-derivatives, but only PA1409 and much more efficiently PA3774 can also process the plain acetylated lysine substrate. P. aeruginosa is able to utilize acetylcadaverine and acetylputrescine as a carbon source under glucose starvation. If either the PA0321 or the PA1409 but not the PA3774 gene is disrupted, the growth of P. aeruginosa is reduced and delayed. In addition, we were able to show that the APAH inhibitors SAHA and SATFMK induce biofilm formation in both PA14 and PAO1 wildtype strains. P. aeruginosa has two functional APAHs, PA0321 and PA1409 which enable the utilization of acetylpolyamines for the metabolism of P. aeruginosa. In contrast, the physiological role of the predicted APAH, PA3774, remains to be elucidated. Its ability to deacetylate synthetic acetylated lysine substrates points to a protein deacetylation functionality with yet unknown substrates. The online version of this article (doi:10.1186/s12858-016-0063-z) contains supplementary material, which is available to authorized users.