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猪链球菌中甲硫氨酸转运蛋白MetQ的鉴定及其对毒力和生物膜形成的作用

Identification of the methionine transporter MetQ in Streptococcus suis and its contribution to virulence and biofilm formation.

作者信息

Bosch Camila, García Carla, Saralegui Luis, van Beek Lucille, de Jonge Marien I, Marín Clara, Arenas Jesús

机构信息

Unit of Microbiology and Immunology, Faculty of Veterinary, University of Zaragoza, Saragossa, Spain.

Agrofood Research University Institute of Aragon (IA2), University of Saragossa - CITA, Zaragoza, Spain.

出版信息

Vet Res. 2025 May 8;56(1):99. doi: 10.1186/s13567-025-01522-y.

Abstract

Streptococcus suis is a Gram-positive bacterium responsible for various infections in both pigs and humans. This study investigates the role of methionine acquisition in the growth and virulence of S. suis. The putative methionine transport system is organised as an operon comprising the metQ gene and genes encoding a transposase and an ATPase, forming a typical tripartite ABC transporter. This operon is conserved across multiple streptococcal species, including both animal and human pathogens. We examined whether MetQ functions as a methionine-binding protein and its role in bacterial infection. Using Western blotting and flow cytometry with a specific antiserum, we demonstrated that MetQ is produced in vitro by the S. suis reference strain P1/7 under methionine-limited conditions and is located on the bacterial cell surface. Growth assays in chemically defined media revealed that a metQ deletion mutant (P1/7∆metQ) exhibited impaired growth under methionine-restricted conditions but grew normally in a nutrient-rich medium, suggesting that MetQ primarily transports methionine. Isothermal Titration Calorimetry demonstrated that MetQ binds L-methionine with a dissociation constant (K) of 7.1 µM. In a murine infection model, the metQ mutant showed reduced dissemination to internal organs compared to the wild type. Furthermore, the mutant showed decreased intracellular survival in murine macrophages and increased sensitivity to oxidative stress, while exhibited enhanced biofilm formation compared to the wild type. Our findings indicate that MetQ is essential for methionine uptake under methionine-restricted conditions, which is critical for bacterial nutrition, immune evasion, and pathogenicity during infection.

摘要

猪链球菌是一种革兰氏阳性细菌,可导致猪和人类发生各种感染。本研究调查了甲硫氨酸摄取在猪链球菌生长和毒力中的作用。推测的甲硫氨酸转运系统组织为一个操纵子,由metQ基因以及编码转座酶和ATP酶的基因组成,形成一个典型的三方ABC转运蛋白。该操纵子在多种链球菌物种中保守,包括动物和人类病原体。我们研究了MetQ是否作为甲硫氨酸结合蛋白发挥作用及其在细菌感染中的作用。使用蛋白质免疫印迹法和特异性抗血清进行流式细胞术,我们证明MetQ在甲硫氨酸限制条件下由猪链球菌参考菌株P1/7在体外产生,并位于细菌细胞表面。在化学成分确定的培养基中进行的生长试验表明,metQ缺失突变体(P1/7∆metQ)在甲硫氨酸限制条件下生长受损,但在营养丰富的培养基中正常生长,这表明MetQ主要转运甲硫氨酸。等温滴定量热法表明,MetQ与L-甲硫氨酸结合的解离常数(K)为7.1µM。在小鼠感染模型中,与野生型相比,metQ突变体向内部器官的扩散减少。此外,该突变体在小鼠巨噬细胞中的细胞内存活率降低,对氧化应激的敏感性增加,而与野生型相比,其生物膜形成增强。我们的研究结果表明,MetQ在甲硫氨酸限制条件下对甲硫氨酸摄取至关重要,这对感染期间的细菌营养、免疫逃避和致病性至关重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7220/12063423/87a554bf5b9a/13567_2025_1522_Fig1_HTML.jpg

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