Article
Biochemistry & Molecular Biology
Ryan J. Shirey, Jonathan R. Hart, BanuPriya Sridharan, Scott J. Novick, Lewis D. Turner, Bin Zhou, Alexander L. Nielsen, Lisa M. Eubanks, Lynn Ueno, Mark S. Hixon, Luke L. Lairson, Timothy P. Spicer, Louis D. Scampavia, Patrick R. Griffin, Peter K. Vogt, Kim D. Janda
Summary: A fluorescent small molecule probe was discovered to detect the oncoprotein MYC, leading to the development of a high-throughput screening assay for MYC inhibitor scaffolds. Screening resulted in molecules with better drug-like properties, with one specific lead showing potent inhibition of oncogenic transformation.
BIOORGANIC & MEDICINAL CHEMISTRY
(2021)
Article
Biochemistry & Molecular Biology
Jacob F. Wardman, Lyann Sim, Jennifer Liu, Teresa A. Howard, Andreas Geissner, Phillip M. Danby, Alisdair B. Boraston, Warren W. Wakarchuk, Stephen G. Withers
Summary: Mucin-type O-glycosylation is an important post-translational modification involved in cellular communication. However, the study of O-glycoproteins and O-glycans is challenging due to the lack of available enzymes for their assembly and degradation. To overcome this, a genetically encoded screening method was developed using engineered Escherichia coli to screen for enzymes that act on O-glycoproteins. The method uses an O-glycosylated fluorescence resonance energy transfer probe as a read-out of glycosylation state and can also assay glycosidases and glycosyltransferases. The methodology has potential for the directed evolution of O-glycopeptidases.
NATURE CHEMICAL BIOLOGY
(2023)
Article
Chemistry, Multidisciplinary
Min Gao, Sun Hyeok Lee, Raj Kumar Das, Haw-Young Kwon, Heon Seok Kim, Young-Tae Chang
Summary: The development of a live B-cell selective small molecule probe CDyB has been achieved through unbiased fluorescence library screening. The molecular target of CDyB is identified as the SLC35C2 transporter. The fluorescence of CDyB and the expression of SLC35C2 are positively correlated with B-cell maturation.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Biochemistry & Molecular Biology
Ji-Dong Shen, Xue Cai, Zhi-Qiang Liu, Yu-Guo Zheng
Summary: In this study, a novel fluorescent probe Rho-IDA-CoII was synthesized and successfully used to label and detect His-tagged proteins. The recombinant Bacillus subtilis strain capable of secreting NHase was screened using this probe, and high NHase activity was achieved.
Article
Multidisciplinary Sciences
Parisa Shiri, Veronica Lai, Tara Zepel, Daniel Griffin, Jonathan Reifman, Sean Clark, Shad Grunert, Lars P. E. Yunker, Sebastian Steiner, Henry Situ, Fan Yang, Paloma L. Prieto, Jason E. Hein
Summary: Solubility screening is essential but labor intensive. By developing a flexible robotic system with computer vision and iterative feedback, researchers can successfully measure solubility in multiple solvents autonomously, saving time and resources.
Review
Biochemical Research Methods
Lihao Fu, Erpeng Guo, Jianzhi Zhang, Keyi Li, Yongcan Chen, Tong Si
Summary: Microbial cell factories play a crucial role in converting renewable feedstocks into chemicals and materials. The lack of predictive modeling has made high-throughput screening essential for microbial strain engineering. Mass spectrometry (MS) has emerged as a powerful tool for label-free detection, offering high sensitivity and chemical specificity. Recent advancements in rapid MS assays have significantly improved screening efficiency for microbial libraries, approaching a rate of one sample per second. However, unique challenges in MS screening of strain libraries need to be addressed for future solutions.
CURRENT OPINION IN BIOTECHNOLOGY
(2022)
Review
Pharmacology & Pharmacy
Tuan Xu, Wei Zheng, Ruili Huang
Summary: In response to the COVID-19 pandemic, a panel of assays has been developed to screen drugs for anti-SARS-CoV-2 activity. The study found that multitarget assays were more accurate and efficient, while target-specific assays were better for investigating compound mechanisms of action. Additionally, strict filtering with counter screens may be more harmful than beneficial in identifying true positives, highlighting the importance of developing novel HTS assays against multiple targets in the SARS-CoV-2 life cycle.
DRUG DISCOVERY TODAY
(2021)
Article
Biochemistry & Molecular Biology
Jorge A. Lerma Romero, Christian Meyners, Andreas Christmann, Lisa M. Reinbold, Anna Charalampidou, Felix Hausch, Harald Kolmar
Summary: This article describes a protein engineering strategy that identified variants with higher affinity binding of conformation-specific FKBP51 selective inhibitors. These variants will provide a better understanding of the conformational flexibility of the FKBP51 binding pocket and may enable the isolation of new selective ligands.
FRONTIERS IN MOLECULAR BIOSCIENCES
(2022)
Article
Biotechnology & Applied Microbiology
You Li, Zhen Qin, Fengli Zhang, Shang-Tian Yang
Summary: Reporter gene assay is developed to detect drugs repressing the transcription of survivin in breast cancer cells, and validated for high throughput screening of drugs targeting survivin expression.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Review
Biochemistry & Molecular Biology
Juan Cheng, Xin Li
Summary: This study reviews various activity-based fluorescent probes used in high-throughput screening assays and emphasizes their structure-related working mechanisms. The study also explores the potential of developing more and better probes to boost hit identification and drug development.
CURRENT MEDICINAL CHEMISTRY
(2022)
Article
Chemistry, Physical
Simon Ladeveze, Paul J. Zurek, Tomasz S. Kaminski, Stephane Emond, Florian Hollfelder
Summary: Enzyme discovery and directed evolution are two contemporary approaches for improving biocatalysis in various fields. Screening campaigns for evolving enzymes rely on the sampled fraction of sequence space, while ultrahigh-throughput screening (uHTS) based on microfluidic systems allows for screening rates >1 kHz. However, screening for carbohydrate-active enzymes (CAZymes) that catalyze valuable reactions is challenging due to difficulties in monitoring released carbohydrates. To overcome these issues, microdroplet assays were designed to make optically inactive carbohydrate products visible by specific cascades. Implementing these assays at the picoliter droplet scale enabled the detection of specific complex carbohydrate degradation products. This advancement in screening technology will facilitate the development of specific and efficient biocatalysts for biovalorization.
Review
Chemistry, Inorganic & Nuclear
Caiping Ding, Tianbing Ren
Summary: Near infrared (NIR) fluorescent probes have advantageous features including low background interference, strong tissue penetration, and minimal tissue damage. This paper provides a summary and discussion on the various types and properties of fluorophores used in the design of NIR fluorescent probes. It also discusses the latest research achievements in detecting and imaging reactive oxygen species, reactive nitrogen, and active sulfur in cells or in vivo. Additionally, a concise overview of the future development of NIR molecular fluorescence probes is proposed based on existing research, offering a reference for current research directions.
COORDINATION CHEMISTRY REVIEWS
(2023)
Article
Biochemistry & Molecular Biology
Cheng Peng, Yu-Hsuan Li, Chao-Wu Yu, Ze-Hua Cheng, Jia-Rong Liu, Jui-Ling Hsu, Ling-Wei Hsin, Chen-Tsung Huang, Hsueh-Fen Juan, Ji-Wang Chern, Yi-Sheng Cheng
Summary: MTH1 is a key enzyme that plays a role in DNA repair, preventing the insertion of incorrect nucleotides and reducing cell apoptosis caused by DNA damage. Through screening for lead compounds, several compounds with MTH1 inhibitory activity were identified, with MI0639 showing the highest effective inhibition. Compound derivatives were synthesized to explore the selectivity and specificity of the action, leading to the discovery of two compounds, MI1020 and MI1024, with nanomolar-range IC50 inhibition abilities.
BIOORGANIC CHEMISTRY
(2021)
Article
Biochemistry & Molecular Biology
Katja F. Hellendahl, Maryke Fehlau, Sebastian Hans, Peter Neubauer, Anke Kurreck
Summary: The study introduces a luciferase-based assay for the detection of NK activity, which was validated in a 96-well plate format. The high-throughput assay showed comparable results to HPLC analysis.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Review
Biochemistry & Molecular Biology
Yuan-Xiang Li, Dong-Tai Xie, Ya-Xi Yang, Zhao Chen, Wu-Yingzheng Guo, Wen-Chao Yang
Summary: Enzymes, as biological catalysts, play a vital role in controlling metabolic reactions. The regulation and amount of enzymes in living cells are indicators of the cell's metabolic status. Small-molecule fluorescent probes are of interest due to their high sensitivity, selectivity, and potential for automated detection. However, designing an ideal fluorescent probe for versatile purposes is challenging. In the future, the focus will be on improving the selectivity, sensitivity, and penetration ability of enzyme-targeting fluorescent probes, as well as coupling them with other imaging molecules/technologies.