以血清白蛋白为信号放大器的荧光探针,用于实时检测溶液、动物细胞线粒体和水稻根中的 HSO3-。

Manzoor Ahmad, Shagun Verma, Nancy Singla, Siloni Singh Bhadwal, Satwinderjeet Kaur, Prabhpreet Singh, Subodh Kumar
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引用次数: 0

摘要

线粒体中含硫氨基酸在酶促氧化过程中会释放出内源性 HSO3-,或亚硫酸盐氧化酶不足会导致亚硫酸盐和硫代硫酸盐在生物液体中积累,影响脑线粒体的线粒体平衡,从而引发与神经系统疾病相关的严重临床症状。红色荧光探针 MGQ 在水中会发生自组装,并显示出聚集诱导的荧光淬灭。在 HSA 和 BSA 存在的情况下,MGQ 在 645 纳米波长处的荧光强度分别增加了 143 倍和 179 倍,但并不能明显区分两种白蛋白。对 MGQ 的详细研究是在有 BSA 存在的情况下进行的。其他酶/蛋白质和氨基酸(如胃蛋白酶、胰蛋白酶、溶菌酶、菠萝蛋白酶、赖氨酸、组氨酸、血红蛋白等)的存在不会影响 MGQ 或 MGQ-BSA 溶液的荧光,表明其对 BSA 具有高选择性。BSA 的检测限为 10 nM。在 PBS 缓冲液中,没有 BSA 的 MGQ 不会与 HSO3- 发生反应,在 1 :1 的乙醇-水混合物中反应迟钝。然而,在 BSA/HSA 的密闭空间中,MGQ 显示出信号放大,在加入 HSO3- 后瞬间发生迈克尔型反应,并在≤1.5 分钟内导致荧光强度的比率变化,在 645 纳米波长处红色荧光减少,在 515 纳米波长处绿色荧光出现。HSO3- 的检测限为 4 nM。
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A fluorescent probe with serum albumin as a signal amplifier for real-time sensing of HSO3- in solution, mitochondria of animal cells and rice roots.

Endogenous release of HSO3- during the enzymatic oxidation of sulfur containing amino acids in mitochondria or insufficiency of sulfite oxidase results in the accumulation of sulfite and thiosulfate in biological fluids affecting mitochondrial homeostasis of brain mitochondria associated with serious clinical symptoms related to neurological disorders. The red fluorescent probe MGQ undergoes self-assembly in water and reveals aggregation induced quenching of fluorescence. MGQ reveals 143-fold and 179-fold increases in fluorescence intensity at 645 nm, respectively, in the presence of HSA and BSA and does not significantly differentiate between two albumins. The detailed studies of MGQ have been performed in the presence of BSA. The presence of other enzymes/proteins and amino acids, viz. pepsin, trypsin, lysozyme, Bromelain, lysine, histidine, hemoglobin, etc., does not affect the fluorescence of MGQ or MGQ-BSA solutions and points to high selectivity towards BSA. The limit of detection for BSA is 10 nM. In PBS buffer, MGQ in the absence of BSA does not react with HSO3- and sluggishly in a 1 : 1 ethanol-water mixture. However, in the confined space of BSA/HSA, MGQ displays a signal amplification, undergoes instantaneous Michael type addition of HSO3- and results in a ratiometric change in fluorescence intensity in ≤1.5 min with the decrease of red fluorescence at 645 nm and emergence of green fluorescence at 515 nm. The LOD for the detection of HSO3- is 4 nM.

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来源期刊
Journal of materials chemistry. B
Journal of materials chemistry. B 化学科学, 工程与材料, 生命科学, 分析化学, 高分子组装与超分子结构, 高分子科学, 免疫生物学, 免疫学, 生化分析及生物传感, 组织工程学, 生物力学与组织工程学, 资源循环科学, 冶金与矿业, 生物医用高分子材料, 有机高分子材料, 金属材料的制备科学与跨学科应用基础, 金属材料, 样品前处理方法与技术, 有机分子功能材料化学, 有机化学
CiteScore
12.00
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0.00%
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审稿时长
1 months
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