Novel flavonoid-based fluorescent probes for site-specific differentiation of human and bovine serum albumin: Visualizing quantitative research in urinary HSA

IF 4.7 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Bioorganic Chemistry Pub Date : 2025-09-13 DOI:10.1016/j.bioorg.2025.108988

Xianfeng Ma , Zixian Shen , Xiaolan Liang , Jiahui Wang , Hua Wang , Mingqi Wang , Xiaofeng Yu

{"title":"Novel flavonoid-based fluorescent probes for site-specific differentiation of human and bovine serum albumin: Visualizing quantitative research in urinary HSA","authors":"Xianfeng Ma , Zixian Shen , Xiaolan Liang , Jiahui Wang , Hua Wang , Mingqi Wang , Xiaofeng Yu","doi":"10.1016/j.bioorg.2025.108988","DOIUrl":null,"url":null,"abstract":"<div><div>Human serum albumin (HSA) is the most abundant protein in serum and is essential for maintaining human health. Abnormal levels have been associated with a variety of diseases, making it an important disease-diagnostic biomarker and therapeutic monitoring indicator. Due to the high structural similarity between human serum albumin (HSA) and bovine serum albumin (BSA), the detection and differentiation of these two proteins have garnered significant attention. In this study, we reported three rationally engineered flavonoid fluorescent probes capable of selectively detecting and distinguishing HSA from BSA. Among these compounds, <strong>HTN2</strong> exhibited a markedly enhanced fluorescence response to HSA with a detection limit of 4.8 nM, while <strong>HTN3</strong> demonstrated a significantly amplified fluorescence response to BSA with a detection limit of 1.4 nM. Notably, the interaction between <strong>HTN2</strong> and HSA was entropy-driven, whereas <strong>HTN3</strong> binding to BSA was enthalpy-driven. Molecular docking experiments investigated the intrinsic reasons for the differing selectivity of the nitrogen-containing six-membered heterocycles toward HSA versus BSA. Furthermore, the <strong>HTN2</strong> probes were successfully applied to detect HSA in real urine samples, offering a reliable approach for HSA analysis in biological fluids. The results showed that introducing the nitrogen heterocycle improved selectivity for HSA and BSA. <strong>HTN2</strong> entered the hydrophobic pocket of HSA effectively; however, when bound to BSA, the nitrogen heterocycle was partially exposed and formed interactions poorly. This provides an effective method for designing specific probes</div></div>","PeriodicalId":257,"journal":{"name":"Bioorganic Chemistry","volume":"165 ","pages":"Article 108988"},"PeriodicalIF":4.7000,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Bioorganic Chemistry","FirstCategoryId":"92","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0045206825008685","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Human serum albumin (HSA) is the most abundant protein in serum and is essential for maintaining human health. Abnormal levels have been associated with a variety of diseases, making it an important disease-diagnostic biomarker and therapeutic monitoring indicator. Due to the high structural similarity between human serum albumin (HSA) and bovine serum albumin (BSA), the detection and differentiation of these two proteins have garnered significant attention. In this study, we reported three rationally engineered flavonoid fluorescent probes capable of selectively detecting and distinguishing HSA from BSA. Among these compounds, HTN2 exhibited a markedly enhanced fluorescence response to HSA with a detection limit of 4.8 nM, while HTN3 demonstrated a significantly amplified fluorescence response to BSA with a detection limit of 1.4 nM. Notably, the interaction between HTN2 and HSA was entropy-driven, whereas HTN3 binding to BSA was enthalpy-driven. Molecular docking experiments investigated the intrinsic reasons for the differing selectivity of the nitrogen-containing six-membered heterocycles toward HSA versus BSA. Furthermore, the HTN2 probes were successfully applied to detect HSA in real urine samples, offering a reliable approach for HSA analysis in biological fluids. The results showed that introducing the nitrogen heterocycle improved selectivity for HSA and BSA. HTN2 entered the hydrophobic pocket of HSA effectively; however, when bound to BSA, the nitrogen heterocycle was partially exposed and formed interactions poorly. This provides an effective method for designing specific probes

Abstract Image

查看原文本刊更多论文

基于黄酮类化合物的新型荧光探针用于人与牛血清白蛋白的位点特异性分化：尿HSA的可视化定量研究

人血清白蛋白（HSA）是血清中含量最多的蛋白质，对维持人体健康至关重要。异常水平与多种疾病相关，是重要的疾病诊断生物标志物和治疗监测指标。由于人血清白蛋白（HSA）和牛血清白蛋白（BSA）在结构上的高度相似性，这两种蛋白的检测和分化受到了广泛的关注。在这项研究中，我们报道了三种合理设计的类黄酮荧光探针，能够选择性地检测和区分HSA和BSA。其中，HTN2对HSA的荧光响应显著增强，检测限为4.8 nM； HTN3对BSA的荧光响应显著增强，检测限为1.4 nM。值得注意的是，HTN2与HSA之间的相互作用是熵驱动的，而HTN3与BSA的结合是焓驱动的。分子对接实验探讨了含氮六元杂环对人血清白蛋白和牛血清白蛋白选择性差异的内在原因。此外，HTN2探针已成功用于检测真实尿液样本中的HSA，为生物体液中的HSA分析提供了可靠的方法。结果表明，氮杂环的引入提高了HSA和BSA的选择性。HTN2有效进入HSA疏水袋；然而，当氮杂环与牛血清白蛋白结合时，氮杂环部分暴露，相互作用形成不良。这为设计特异性探针提供了有效的方法

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Bioorganic Chemistry 生物-生化与分子生物学

CiteScore

9.70

自引率

3.90%

发文量

679

审稿时长

31 days

期刊介绍： Bioorganic Chemistry publishes research that addresses biological questions at the molecular level, using organic chemistry and principles of physical organic chemistry. The scope of the journal covers a range of topics at the organic chemistry-biology interface, including: enzyme catalysis, biotransformation and enzyme inhibition; nucleic acids chemistry; medicinal chemistry; natural product chemistry, natural product synthesis and natural product biosynthesis; antimicrobial agents; lipid and peptide chemistry; biophysical chemistry; biological probes; bio-orthogonal chemistry and biomimetic chemistry. For manuscripts dealing with synthetic bioactive compounds, the Journal requires that the molecular target of the compounds described must be known, and must be demonstrated experimentally in the manuscript. For studies involving natural products, if the molecular target is unknown, some data beyond simple cell-based toxicity studies to provide insight into the mechanism of action is required. Studies supported by molecular docking are welcome, but must be supported by experimental data. The Journal does not consider manuscripts that are purely theoretical or computational in nature. The Journal publishes regular articles, short communications and reviews. Reviews are normally invited by Editors or Editorial Board members. Authors of unsolicited reviews should first contact an Editor or Editorial Board member to determine whether the proposed article is within the scope of the Journal.