Academic Journal
A Live-Cell Fluorescence High-Throughput Screening Method for Antimicrobial Peptides Based on the SpyTag/SpyCatcher System.
| Τίτλος: | A Live-Cell Fluorescence High-Throughput Screening Method for Antimicrobial Peptides Based on the SpyTag/SpyCatcher System. |
|---|---|
| Συγγραφείς: | Li J; Key Laboratory of Geriatric Nutrition and Health (Beijing Technology and Business University), Ministry of Education, Beijing 100048, China.; School of Light Industry Science and Engineering, Beijing Technology and Business University (BTBU), Beijing 100048, China., Wen S; Key Laboratory of Geriatric Nutrition and Health (Beijing Technology and Business University), Ministry of Education, Beijing 100048, China.; School of Light Industry Science and Engineering, Beijing Technology and Business University (BTBU), Beijing 100048, China., Wang F; Key Laboratory of Geriatric Nutrition and Health (Beijing Technology and Business University), Ministry of Education, Beijing 100048, China.; School of Light Industry Science and Engineering, Beijing Technology and Business University (BTBU), Beijing 100048, China. |
| Πηγή: | Journal of agricultural and food chemistry [J Agric Food Chem] 2026 Apr 22; Vol. 74 (15), pp. 12261-12270. Date of Electronic Publication: 2026 Apr 14. |
| Τύπος έκδοσης: | Journal Article; Evaluation Study |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 0374755 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-5118 (Electronic) Linking ISSN: 00218561 NLM ISO Abbreviation: J Agric Food Chem Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: Washington, American Chemical Society. |
| Ιατρικοί όροι (MeSH): | High-Throughput Screening Assays*/methods , High-Throughput Screening Assays*/instrumentation , Antimicrobial Peptides*/pharmacology , Antimicrobial Peptides*/chemistry , Fluorescence Resonance Energy Transfer*/methods , Anti-Bacterial Agents*/pharmacology , Anti-Bacterial Agents*/chemistry , Antimicrobial Cationic Peptides*/pharmacology , Antimicrobial Cationic Peptides*/chemistry, Escherichia coli/drug effects ; Escherichia coli/growth & development ; Bacillus subtilis/drug effects ; Microbial Sensitivity Tests ; Fluorescence |
| Περίληψη: | Antimicrobial peptides (AMPs) are key to innate immunity but face challenges in rational design due to their diversity and complex mechanisms. The lack of sensitive and full function-based high-throughput screening methods for AMP variants has further impeded systematic exploration and optimization. To address this, we developed a high-throughput screening method for AMPs integrating SpyTag/SpyCatcher click biology and fluorescence resonance energy transfer (FRET). Attributing to the ultrahigh reaction rate and specificity of the SpyTag/SpyCatcher pair, this screening method allows real-time fluorometric detection of the cell lytic activity of AMPs against their natural targets, i.e., bacterial cells. Applied to an indolicidin proline-scanning library, this approach identified variants with enhanced activity against E. coli and B. subtilis. Collectively, this work establishes a function-based, high-throughput screening platform for AMPs that can fully reflect their functional complexity, thereby providing an efficient and scalable tool for screening AMP libraries and facilitating data-driven optimization and functional analysis. |
| Contributed Indexing: | Keywords: FRET; SpyTag/SpyCatcher; antimicrobial peptides; high-throughput screening |
| Substance Nomenclature: | 0 (Antimicrobial Peptides) 0 (Anti-Bacterial Agents) 0 (Antimicrobial Cationic Peptides) |
| Entry Date(s): | Date Created: 20260414 Date Completed: 20260715 Latest Revision: 20260715 |
| Update Code: | 20260715 |
| DOI: | 10.1021/acs.jafc.5c17550 |
| PMID: | 41978950 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 1520-5118 |
|---|---|
| DOI: | 10.1021/acs.jafc.5c17550 |