Unité Pathogenèse de Helicobacter, Département de Microbiologie, UMR CNRS 6047, Institut Pasteur, Université Paris Cité , Paris, France.
Université Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur , Paris, France.
mBio. 2023 Oct 31;14(5):e0096723. doi: 10.1128/mbio.00967-23. Epub 2023 Aug 16.
Correct folding of proteins represents a crucial step for their functions. Among the chaperones that control protein folding, the ubiquitous PPIases catalyze the /-isomerization of peptidyl-prolyl bonds. Only few protein targets of PPIases have been reported in bacteria. To fill this knowledge gap, we performed a large-scale two-hybrid screen to search for targets of the and SlyD PPIase-metallochaperone. SlyD from both organisms interacts with enzymes (i) containing metal cofactors, (ii) from the central metabolism tricarboxylic acid (TCA) cycle, and (iii) involved in the formation of the essential and ancestral Fe-S cluster cofactor. and mutants present similar phenotypes of diminished susceptibility to antibiotics and to oxidative stress. In , measurements of the intracellular ATP content, proton motive force, and activity of TCA cycle proteins suggest that SlyD regulates TCA cycle enzymes by controlling the formation of their indispensable Fe-S clusters.
蛋白质的正确折叠是其发挥功能的关键步骤。在控制蛋白质折叠的伴侣分子中，普遍存在的 PPIases 催化肽基脯氨酰键的/-异构化。在细菌中，只有少数 PPIases 的蛋白质靶标被报道。为了填补这一知识空白，我们进行了大规模的双杂交筛选，以寻找 和 SlyD PPIase-金属伴侣的靶标。两种生物体的 SlyD 均与以下酶相互作用：（i）含有金属辅因子的酶，（ii）来自中心代谢三羧酸 (TCA) 循环的酶，以及（iii）参与必需和原始 Fe-S 簇辅因子形成的酶。 和 突变体表现出对抗生素和氧化应激敏感性降低的相似表型。在 中，对细胞内 ATP 含量、质子动力势和 TCA 循环蛋白活性的测量表明，SlyD 通过控制其不可或缺的 Fe-S 簇的形成来调节 TCA 循环酶。
