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Enhancing Peptide Cyclization: Unveiling the Key Roles of Coupling Reagents and Heat for Optimal Results 增强肽环化:揭示偶联试剂和热对最佳结果的关键作用。
IF 2.6 4区 生物学
ChemBioChem Pub Date : 2025-06-04 DOI: 10.1002/cbic.202500375
Kim Gaudin, Alexie Bobe, Vijay Kumar Pal, Céline Corcelle, Kelton Rodrigues Souza, Cécilia Ménard-Moyon
{"title":"Enhancing Peptide Cyclization: Unveiling the Key Roles of Coupling Reagents and Heat for Optimal Results","authors":"Kim Gaudin,&nbsp;Alexie Bobe,&nbsp;Vijay Kumar Pal,&nbsp;Céline Corcelle,&nbsp;Kelton Rodrigues Souza,&nbsp;Cécilia Ménard-Moyon","doi":"10.1002/cbic.202500375","DOIUrl":"10.1002/cbic.202500375","url":null,"abstract":"<p>Cyclic peptides offer several advantages over their linear counterparts, including enhanced structural stability due to their rigid conformation and increased resistance to enzymatic proteolysis. Additionally, their ring structure and constrained conformation reduce the entropic cost upon binding to receptors and other biological targets, leading to higher binding affinity and specificity. However, peptide macrocyclization is often synthetically challenging due to factors such as reduced entropy, oligomer formation, and <i>C</i>-terminal epimerization. The conventional approach for synthesizing cyclic peptides involves the direct coupling of amine and carboxyl termini in the solution phase, using protected side-chain peptides and coupling reagents. Despite this, improving the efficiency of head-to-tail cyclization remains a key challenge. In this study, the cyclization of a <i>de novo</i> octapeptide composed of alternating <span>l</span>- and <span>d</span>-amino acids is optimized, in which the impact of various factors on the cyclization process is examined, including coupling reagents, temperature, heating method, chaotropic agents, solvent, and concentration. This investigation has not only led to the identification of efficient cyclization conditions, but also provides a valuable framework for the cyclization of other challenging peptide sequences. The insights gained in this study contribute to the field of peptide chemistry, expanding the understanding of peptide cyclization reactions.</p>","PeriodicalId":140,"journal":{"name":"ChemBioChem","volume":"26 14","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144214467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Catalytic Characterization of α-Regioselective Halohydrin Dehalogenase from Novosphingobium resinovorum 新藻α-区域选择性卤代醇脱卤酶的催化表征。
IF 2.6 4区 生物学
ChemBioChem Pub Date : 2025-06-04 DOI: 10.1002/cbic.202500163
Hui-Hui Wang, Miao An, Xin-Yu Wang, Yang Zhao, Rui-Ling Yan, Nan-Wei Wan
{"title":"Catalytic Characterization of α-Regioselective Halohydrin Dehalogenase from Novosphingobium resinovorum","authors":"Hui-Hui Wang,&nbsp;Miao An,&nbsp;Xin-Yu Wang,&nbsp;Yang Zhao,&nbsp;Rui-Ling Yan,&nbsp;Nan-Wei Wan","doi":"10.1002/cbic.202500163","DOIUrl":"10.1002/cbic.202500163","url":null,"abstract":"<p>Halohydrin dehalogenases are highly valuable biocatalysts for chiral compound synthesis due to their diverse catalytic capabilities and inherent stereoselectivity. Historically, research on halohydrin dehalogenases is confined to a limited number of enzymes. However, with the advent of advanced gene mining methods, a growing number of halohydrin dehalogenases have been discovered and characterized, revealing a diverse array of new structures and catalytic functions. While halohydrin dehalogenases predominantly exhibit <i>β</i>-regioselectivity in the ring-opening of styrene oxides, a few α-regioselective halohydrin dehalogenases have also been identified. In this study, a new α-regioselective halohydrin dehalogenase, HHDHnsr, is purified and analyzed, and its application in the azide-mediated α-regioselective azidolysis of styrene oxide is explored. The findings highlight the potential of HHDHnsr as α-regioselective biocatalyst for the asymmetric synthesis of <i>β</i>-azidoalcohols, offering valuable insights for the future identification of α-regioselective HHDHs in enzyme mining.</p>","PeriodicalId":140,"journal":{"name":"ChemBioChem","volume":"26 13","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144223812","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Mammalian Tolerance to Amino Acid Heterochirality 哺乳动物对氨基酸杂手性的耐受性。
IF 2.6 4区 生物学
ChemBioChem Pub Date : 2025-06-03 DOI: 10.1002/cbic.202500273
Sakiko Taniguchi, Kenichiro Adachi, Xuan Tran, Masataka Suzuki, Jumpei Sasabe
{"title":"Mammalian Tolerance to Amino Acid Heterochirality","authors":"Sakiko Taniguchi,&nbsp;Kenichiro Adachi,&nbsp;Xuan Tran,&nbsp;Masataka Suzuki,&nbsp;Jumpei Sasabe","doi":"10.1002/cbic.202500273","DOIUrl":"10.1002/cbic.202500273","url":null,"abstract":"<p>Organisms use amino acids predominantly in <span>l-</span>configuration. In contrast, a series of studies show that a variety of <span>d-</span>amino acids also occur in mammals, and amino acid homochirality is not complete. Mammals de novo synthesize most amino acids with <span>l-</span>configuration, but serine and aspartate are converted from <span>l-</span> to <span>d-</span>configuration by endogenous enzymes. In addition to endogenous syntheses of <span>d-</span>amino acids, symbiotic bacteria in mammals chiral-convert amino acids, including alanine, glutamate, proline, and leucine in the intestine, creating a heterochiral inner environment. <span>d-</span>amino acids are distributed in distinctive patterns among organs and have physiological roles in the central nervous, endocrine, and immune systems. Mammals manage such diverse <span>d-</span>amino acids with catabolism and excretion into urine at individual levels. In contrast, at the cellular levels an enantioselection mechanism to regulate chiral homeostasis of amino acids has remained unclear. In protein synthesis, the ribosome has a sophisticated system to eliminate <span>d-</span>amino acids, whereas non-ribosomal synthesis also utilizes <span>d-</span>amino acids. Furthermore, amino acid residues in proteins/peptides can be isomerized post-translationally through enzymatic or spontaneous processes. This manuscript overviews how the chiral balance of free amino acids or residues in proteins is maintained in mammals at the individual and cellular levels.</p>","PeriodicalId":140,"journal":{"name":"ChemBioChem","volume":"26 13","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cbic.202500273","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144214468","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Engineering Glucagon via Molecular and Formulation Strategies: From Natural Hormone to Effective and Stable Therapeutics. 通过分子和配方策略的工程胰高血糖素:从天然激素到有效和稳定的治疗。
IF 2.6 4区 生物学
ChemBioChem Pub Date : 2025-06-03 DOI: 10.1002/cbic.202500270
Xin Li, Yani Wang, Zican Chen, Zhongping Tan
{"title":"Engineering Glucagon via Molecular and Formulation Strategies: From Natural Hormone to Effective and Stable Therapeutics.","authors":"Xin Li, Yani Wang, Zican Chen, Zhongping Tan","doi":"10.1002/cbic.202500270","DOIUrl":"https://doi.org/10.1002/cbic.202500270","url":null,"abstract":"<p><p>Glucagon, a 29-amino acid pancreatic hormone, plays a central role in glucose homeostasis through activation of the glucagon receptor (GCGR). While clinically essential for hypoglycemia rescue, its broader therapeutic applications face limitations due to challenging biophysical properties including poor solubility, strong aggregation tendency, and chemical instability, which currently require lyophilized formulations. Recent advances in peptide engineering and formulation science have enabled the development of next-generation glucagon analogs with enhanced stability and ready-to-use profiles. Moving beyond its classical role in glucose elevation, glucagon is now recognized as a multifunctional metabolic regulator influencing lipid metabolism, energy expenditure, and cardiorenal function. This expanded understanding has driven the development of innovative GCGR-targeting therapies including multi-receptor agonists such as glucagon-like peptide-1 receptor (GLP-1R)/GCGR co-agonists for metabolic disorders and advanced dual-hormone delivery systems. This review examines the evolving pharmacology of glucagon by analyzing: (1) structural optimization and formulation strategies to address physicochemical challenges, (2) innovative delivery technologies, and (3) emerging therapeutic applications in metabolic diseases. Through critical assessment of translational challenges, this work bridges molecular insights with clinical innovation in glucagon-based therapeutics.</p>","PeriodicalId":140,"journal":{"name":"ChemBioChem","volume":" ","pages":"e202500270"},"PeriodicalIF":2.6,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144207257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Recent Advances in Structure, Biofunction, Detection, and Disease Therapeutic Targeting of Cytochrome P450 细胞色素P450的结构、生物功能、检测及疾病治疗靶点研究进展。
IF 2.6 4区 生物学
ChemBioChem Pub Date : 2025-05-31 DOI: 10.1002/cbic.202500278
Shuo Yang, Lan Wang, Jiayi Wu, Tianwei Liu, Siying Pei, Qian Zhou, Qiong Wu
{"title":"Recent Advances in Structure, Biofunction, Detection, and Disease Therapeutic Targeting of Cytochrome P450","authors":"Shuo Yang,&nbsp;Lan Wang,&nbsp;Jiayi Wu,&nbsp;Tianwei Liu,&nbsp;Siying Pei,&nbsp;Qian Zhou,&nbsp;Qiong Wu","doi":"10.1002/cbic.202500278","DOIUrl":"10.1002/cbic.202500278","url":null,"abstract":"<p>The heme monooxygenase cytochrome P450 (CYPs), which play a major role in human drug metabolism, is predominantly found in the liver and intestines. CYPs are essential for the metabolism of therapeutic drugs and the treatment of diseases. It catalyzes the oxidative metabolism of several endogenous and exogenous chemicals. The expression and biological functions of CYP are affected by environmental factors, genetic variations, and physiological conditions, which can lead to adverse effects and reduce the therapeutic efficacy of drugs in humans. Therefore, there has been considerable interest in the development of probes that can detect the activity of CYPs in complex living systems. This review explores the recent advancements in our understanding of the structure and function of CYPs. It highlights the need to develop specific probes for various CYP subtypes using nonoptical, fluorescent, and bioluminescent substrates. Furthermore, it summarizes the potential of certain CYP subtypes as therapeutic targets for various diseases. Finally, the challenges and prospects of probes for monitoring CYP activity are explored.</p>","PeriodicalId":140,"journal":{"name":"ChemBioChem","volume":"26 14","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144191303","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Comparative Function Analysis of Self-Amplifying mRNA and Self-Amplifying DNA 自扩增mRNA与自扩增DNA的功能比较分析。
IF 2.6 4区 生物学
ChemBioChem Pub Date : 2025-05-30 DOI: 10.1002/cbic.202500110
Wenting Li, Yiming Wang, Chen Wang, Yi-Xin Huo, Yuan Lu
{"title":"Comparative Function Analysis of Self-Amplifying mRNA and Self-Amplifying DNA","authors":"Wenting Li,&nbsp;Yiming Wang,&nbsp;Chen Wang,&nbsp;Yi-Xin Huo,&nbsp;Yuan Lu","doi":"10.1002/cbic.202500110","DOIUrl":"10.1002/cbic.202500110","url":null,"abstract":"<p>A promising approach to address the transient translation mRNA drugs is the development of self-amplifying mRNA (SAM). Similarly, SAM's DNA template, self-amplifying DNA (SAD), can also play a role in self-amplifying the target mRNA. However, there is still a lack of sufficient valid information on the comparative analysis of SAM and SAD. Therefore, various vectors and delivery systems are selected to compare and analyze the transfection efficiency of SAM and SAD, and transfection conditions for SAM are optimized. It is found that SAD and SAM have obviously different preferences in transfection conditions, and there is also a gap in their transfection efficiencies under the same conditions. Furthermore, the optimal transfection dose of SAD and SAM and the optimal incubation time after transfection are also determined. This study provides valuable information on SAM transfection and its difference with SAD, and offers an option of selecting appropriate nucleic acid vectors for drug development.</p>","PeriodicalId":140,"journal":{"name":"ChemBioChem","volume":"26 13","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144186107","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A De Novo Designed Metalloprotein Displays Variable Thermal Stability and Binding Stoichiometry with Transition Metal Ions 一种全新设计的金属蛋白具有可变的热稳定性和与过渡金属离子的结合化学计量学。
IF 2.6 4区 生物学
ChemBioChem Pub Date : 2025-05-28 DOI: 10.1002/cbic.202500322
Britt Rooijakkers, Gaya Verhagen, Anneloes Cramer-Blok, Ed Zuidinga, Aimee L. Boyle
{"title":"A De Novo Designed Metalloprotein Displays Variable Thermal Stability and Binding Stoichiometry with Transition Metal Ions","authors":"Britt Rooijakkers,&nbsp;Gaya Verhagen,&nbsp;Anneloes Cramer-Blok,&nbsp;Ed Zuidinga,&nbsp;Aimee L. Boyle","doi":"10.1002/cbic.202500322","DOIUrl":"10.1002/cbic.202500322","url":null,"abstract":"<p>Metal-binding selectivity in natural proteins is determined by multiple factors such as the protein's structure, metal concentration within cellular compartments, and the presence of metallochaperones. The in vitro selectivity of proteins for transition metal ions is largely governed by the Irving–Williams series, which states protein-metal complex stability follows the order Co(II) &lt; Ni(II) &lt; Cu(II) &gt; Zn(II). A de novo protein has been designed that folds in the presence of certain transition metal ions into a monomeric α-helical bundle, with the least stable protein-metal complex being formed with Cu(II). Moreover, when increasing the metal concentration of Cu(II) or Zn(II), more metal ions are incorporated into the protein accompanied by a concurrent decrease in the amount of secondary structure. One reason may be that there is a balance between stability conferred by the coordination of the metal ion(s) and stability conferred by hydrophobic packing of the α-helical bundle. Metals may therefore adopt distorted coordination geometries, or binding of multiple ions may cause distortion of the protein backbone, leading to compromised folding of the protein scaffold, or variable thermal stabilities of the metalloprotein complexes. This protein scaffold therefore contributes to the deciphering of design rules for metal selectivity in proteins.</p>","PeriodicalId":140,"journal":{"name":"ChemBioChem","volume":"26 14","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cbic.202500322","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144172250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Front Cover: Wireframe DNA Origami Capable of Vertex-protruding Transformation (ChemBioChem 10/2025) 封面:线框DNA折纸能够顶点突出转换(ChemBioChem 10/2025)
IF 2.6 4区 生物学
ChemBioChem Pub Date : 2025-05-28 DOI: 10.1002/cbic.202581001
Yosuke Ochi, Wataru Kato, Yoichi Tsutsui, Dr. Yuki Gomibuchi, Daichi Tominaga, Keisuke Sakai, Takeshi Araki, Suzunosuke Yoshitake, Prof. Dr. Takuo Yasunaga, Prof. Dr. Yusuke V. Morimoto, Prof. Dr. Kazuhiro Maeda, Prof. Dr. Junichi Taira, Prof. Dr. Yusuke Sato
{"title":"Front Cover: Wireframe DNA Origami Capable of Vertex-protruding Transformation (ChemBioChem 10/2025)","authors":"Yosuke Ochi,&nbsp;Wataru Kato,&nbsp;Yoichi Tsutsui,&nbsp;Dr. Yuki Gomibuchi,&nbsp;Daichi Tominaga,&nbsp;Keisuke Sakai,&nbsp;Takeshi Araki,&nbsp;Suzunosuke Yoshitake,&nbsp;Prof. Dr. Takuo Yasunaga,&nbsp;Prof. Dr. Yusuke V. Morimoto,&nbsp;Prof. Dr. Kazuhiro Maeda,&nbsp;Prof. Dr. Junichi Taira,&nbsp;Prof. Dr. Yusuke Sato","doi":"10.1002/cbic.202581001","DOIUrl":"https://doi.org/10.1002/cbic.202581001","url":null,"abstract":"<p>The cover picture illustrates the reversible transformation of wireframe polyhedral DNA origami, transitioning between an open-form and a closed-form with protruding vertices. Spacer strands between vertices and edges introduce vertex-specific flexibility, enabling designed vertex protrusion. The transformation is driven by DNA hybridization and strand displacement reactions, allowing the structure to switch reversibly between its open and closed states. More details can be found in article 10.1002/cbic.202401071 by Yusuke Sato and co-workers.\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure>\u0000 </p>","PeriodicalId":140,"journal":{"name":"ChemBioChem","volume":"26 10","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cbic.202581001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144148591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Synthesis and Evaluation of Trehalose-Based Mertansine Warheads for Bacillus Calmette-Guérin Delivery of Anticancer Agents. 用于BCG抗癌药物递送的海藻糖Mertansine战斗部的合成与评价。
IF 2.6 4区 生物学
ChemBioChem Pub Date : 2025-05-27 DOI: 10.1002/cbic.202500390
Michael Grimmeisen, Xuan Wang, Melissa Weldle, Kerstin Sartory, Sara Benkhelifa, Yu Zhang, Trinh Dao, Jonas Meyer, Oliver Gorka, Olaf Groß, Claudia Jessen-Trefzer
{"title":"Synthesis and Evaluation of Trehalose-Based Mertansine Warheads for Bacillus Calmette-Guérin Delivery of Anticancer Agents.","authors":"Michael Grimmeisen, Xuan Wang, Melissa Weldle, Kerstin Sartory, Sara Benkhelifa, Yu Zhang, Trinh Dao, Jonas Meyer, Oliver Gorka, Olaf Groß, Claudia Jessen-Trefzer","doi":"10.1002/cbic.202500390","DOIUrl":"10.1002/cbic.202500390","url":null,"abstract":"<p><p>Nonmuscle invasive bladder cancer (NMIBC) accounts for 75% of bladder cancer cases, with Bacillus Calmette-Guérin (BCG) immunotherapy as the gold standard for high-risk patients. BCG elicits a robust immune response but is limited by adverse effects and resistance. To enhance its efficacy, a trehalose-based conjugation strategy is developed, tethering a cytostatic agent to BCG via a cleavable disulfide linker. This system enables selective drug integration into the BCG envelope and controlled release in tumor cells, aiming to improve therapeutic precision while minimizing toxicity. This approach combines immunotherapy with targeted chemotherapy, offering a promising strategy for NMIBC treatment.</p>","PeriodicalId":140,"journal":{"name":"ChemBioChem","volume":" ","pages":"e2500390"},"PeriodicalIF":2.6,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144149018","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Macromolecular High-Affinity Binding Probed by Advanced Fluorescence Techniques. 先进荧光技术探测大分子高亲和力结合。
IF 2.6 4区 生物学
ChemBioChem Pub Date : 2025-05-27 DOI: 10.1002/cbic.202500283
Alida Meyer, Benno Schedler, Jörg Fitter
{"title":"Macromolecular High-Affinity Binding Probed by Advanced Fluorescence Techniques.","authors":"Alida Meyer, Benno Schedler, Jörg Fitter","doi":"10.1002/cbic.202500283","DOIUrl":"10.1002/cbic.202500283","url":null,"abstract":"<p><p>Due to the extreme sensitivity and the intrinsic selectivity of fluorescence techniques, high-affinity binding can be measured even at extremely low molecule concentrations in the picomolar range. In particular, modern advanced techniques with fluorescence microscopes have provided considerable methodological advancements in recent years. Here, a brief description of the basic physical principles of fluorescence detection and its experimental measurement setups are provided. For interacting biomolecules in solution, confocal fluorescence microscopy enables some very effective approaches to characterize binding in complex sample environments and with small sample consumption. In addition to standard techniques with bulk samples in classical spectrometers, applications with single-molecule Förster resonance energy transfer, two-color coincidence detection, and fluorescence correlation spectroscopy are presented. The strength of the more advanced techniques lies in their broad applicability, ranging from fluorescence-based genetically encoded biosensors for use in living cells to the high controllability in the measurement of binding curves even at very low molecule concentrations. The advantages and limitations of the individual techniques are compared and recent state-of-the-art applications are discussed.</p>","PeriodicalId":140,"journal":{"name":"ChemBioChem","volume":" ","pages":"e2500283"},"PeriodicalIF":2.6,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144149003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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