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研究来自Pseudomonas aeruginosa的异酸酶 (ICL) 的结构功能中的外围领域

Alma Laura Díaz-Pérez¹, César Díaz-Pérez², Roxana Yughadi Gaona-García¹

¹Lab. de Biotecnología Microbiana, Instituto de Investigaciones Químico Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Edif. U-3, Ciudad Universitaria, 58030, Morelia, Mich., Mexico.

World journal of microbiology & biotechnology

|October 11, 2023

概括

Pseudomonas aeruginosa异酸酶 (PaICL) 需要域II和扩展的动机来实现功能和稳定性. 对于PaICL来说,C端是非常重要的.

科学领域:

微生物学微生物学
酶学是一种酶学.
细菌病原体的产生

背景情况:

Pseudomonas aeruginosa 的营养同化是殖民和致病性的关键.
异酸盐酶 (ICL) 通过氧酸盐循环促进碳同化,作为一种毒性因子.
属于子家族3的PaICL具有独特的结构特征,如域II和扩展图案.

研究的目的:

为了研究PaICL的域II,扩展图案和N/C-termini的功能作用.
确定这些区域在PaICL酶活性和P. aeruginosa毒性中的参与.
阐明PaICL功能的结构基础及其对病毒毒性的贡献.

主要方法:

位点定向的突变发生,以删除域II和扩展动机.
构建和描述N端和C端的His6标记的PaICL变体.
在ICL突变菌株中进行酶活性测定和补充研究.
在P. aeruginosa PAO1模型中评估细菌毒性.

主要成果:

删除域II和扩展图案使PaICL失活,导致结构不稳定.
与C端变体相比,N端His6标记的PaICL在酶上更有效,结构上更稳定.
鉴定出C末端对PaICL四分体结构构造至关重要.
与N端和C端标记PaICL的补充恢复了ICL突变体中的毒性.

结论:

域II和扩展图案对于PaICL结构和功能至关重要.
在维护PaICL的四分体结构方面,C端起着至关重要的作用.
在P. aeruginosa中,PaICL对于酸盐同化和毒性是不可或缺的.

关键词:

细菌病原发生学异酸盐溶解酶是什么意思蛋白质结构蛋白质结构伪omonas aeruginosa 这种类型的病毒.一个TIM-桶.

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