Structural and Kinetic Characterization of the SpeG Spermidine/Spermine N-acetyltransferase from Methicillin-Resistant Staphylococcus aureus USA300.
Structural and Kinetic Characterization of the SpeG Spermidine/Spermine N-acetyltransferase from Methicillin-Resistant Staphylococcus aureus USA300.
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耐甲氧西林金黄色葡萄球菌USA300的SpeG亚精胺/精胺N-乙酰转移酶的结构和动力学特征
DOI：
10.3390/cells12141829
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发表时间：
2023-07-12
期刊：
影响因子：
6
通讯作者：
Forwood, Jade K. K.
中科院分区：
文献类型：
--
作者：
Tsimbalyuk, Sofiya;Shornikov, Aleksander;Srivastava, Parul;Le, Van Thi Bich;Warren, Imani;Khandokar, Yogesh B. B.;Kuhn, Misty L. L.;Forwood, Jade K. K.
关键词：
多胺是一种简单而关键的分子，在许多致病性和非致病性生物体中具有不同的作用。调节多胺浓度影响对细胞生长、应激和毒性重要的基因和蛋白质的转录和翻译。多胺浓度在细胞内维持的一种方式是通过亚精胺/精胺N-乙酰基转移酶（SSAT）使细胞内多胺乙酰化，从而使它们可以输出。细菌SpeG酶是一种SSAT，与先前已表征的其他SSAT相比，其表现出独特的十二聚体结构和变构位点。虽然它的整体3D结构是保守的，但它在不同细菌病原体中的存在和作用并不一致。例如，并非所有细菌的基因组中都有speG编码;在一些细菌中，speG基因存在，但已经沉默，而在其他细菌中，它已经在移动的遗传元件上获得。后者是耐甲氧西林金黄色葡萄球菌（MRSA）USA 300的情况，它似乎有助于发病机制。为了更好地了解MRSA USA 300菌株（SaSpeG）SpeG的结构/功能关系，我们在精胺存在和不存在的情况下测定了其X射线晶体结构。此外，我们发现SaSpeG的寡聚状态是动态的，其均一性受多胺和AcCoA的影响。酶动力学分析表明，多胺预孵育显着影响对精胺和亚精胺的正协同作用和酶的催化效率。此外，我们表明细菌SpeG酶不具有乙酰化亚精胺的氨基丙基与氨基丁基末端的等效能力。总的来说，这项研究提供了新的见解，将有助于了解SpeG酶及其在分子水平上对致病性和非致病性细菌的作用。
Polyamines are simple yet critical molecules with diverse roles in numerous pathogenic and non-pathogenic organisms. Regulating polyamine concentrations affects the transcription and translation of genes and proteins important for cell growth, stress, and toxicity. One way polyamine concentrations are maintained within the cell is via spermidine/spermine N-acetyltransferases (SSATs) that acetylate intracellular polyamines so they can be exported. The bacterial SpeG enzyme is an SSAT that exhibits a unique dodecameric structure and allosteric site compared to other SSATs that have been previously characterized. While its overall 3D structure is conserved, its presence and role in different bacterial pathogens are inconsistent. For example, not all bacteria have speG encoded in their genomes; in some bacteria, the speG gene is present but has become silenced, and in other bacteria, it has been acquired on mobile genetic elements. The latter is the case for methicillin-resistant Staphylococcus aureus (MRSA) USA300, where it appears to aid pathogenesis. To gain a greater understanding of the structure/function relationship of SpeG from the MRSA USA300 strain (SaSpeG), we determined its X-ray crystal structure in the presence and absence of spermine. Additionally, we showed the oligomeric state of SaSpeG is dynamic, and its homogeneity is affected by polyamines and AcCoA. Enzyme kinetic assays showed that pre-incubation with polyamines significantly affected the positive cooperativity toward spermine and spermidine and the catalytic efficiency of the enzyme. Furthermore, we showed bacterial SpeG enzymes do not have equivalent capabilities to acetylate aminopropyl versus aminbutyl ends of spermidine. Overall, this study provides new insight that will assist in understanding the SpeG enzyme and its role in pathogenic and non-pathogenic bacteria at a molecular level.
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DOI：
10.1002/pol.1980.130180406
发表时间：
1980-01-01
期刊：
JOURNAL OF POLYMER SCIENCE PART C-POLYMER LETTERS
影响因子：
--
作者：
HAGLUND, AC;MARSDEN, NVB
通讯作者：
MARSDEN, NVB
DOI：
10.1107/s2059798319006545
发表时间：
2019-06-01
影响因子：
2.2
作者：
Filippova, Ekaterina V.;Weigand, Steven;Anderson, Wayne F.
通讯作者：
Anderson, Wayne F.
影响因子：
8
作者：
Majorek, Karolina A.;Kuhn, Misty L.;Minor, Wladek
通讯作者：
Minor, Wladek
影响因子：
6.4
作者：
Diep, Binh An;Stone, Gregory G.;Chambers, Henry F.
通讯作者：
Chambers, Henry F.
影响因子：
14.9
作者：
CORPET, F
通讯作者：
CORPET, F