Multivalent binding kinetics resolved by fluorescence proximity sensing
Please always quote using this URN: urn:nbn:de:bvb:20-opus-301157
- Multivalent protein interactors are an attractive modality for probing protein function and exploring novel pharmaceutical strategies. The throughput and precision of state-of-the-art methodologies and workflows for the effective development of multivalent binders is currently limited by surface immobilization, fluorescent labelling and sample consumption. Using the gephyrin protein, the master regulator of the inhibitory synapse, as benchmark, we exemplify the application of Fluorescence proximity sensing (FPS) for the systematic kinetic andMultivalent protein interactors are an attractive modality for probing protein function and exploring novel pharmaceutical strategies. The throughput and precision of state-of-the-art methodologies and workflows for the effective development of multivalent binders is currently limited by surface immobilization, fluorescent labelling and sample consumption. Using the gephyrin protein, the master regulator of the inhibitory synapse, as benchmark, we exemplify the application of Fluorescence proximity sensing (FPS) for the systematic kinetic and thermodynamic optimization of multivalent peptide architectures. High throughput synthesis of +100 peptides with varying combinatorial dimeric, tetrameric, and octameric architectures combined with direct FPS measurements resolved on-rates, off-rates, and dissociation constants with high accuracy and low sample consumption compared to three complementary technologies. The dataset and its machine learning-based analysis deciphered the relationship of specific architectural features and binding kinetics and thereby identified binders with unprecedented protein inhibition capacity; thus, highlighting the value of FPS for the rational engineering of multivalent inhibitors.…
Author: | Clemens Schulte, Alice Soldà, Sebastian Spänig, Nathan Adams, Ivana Bekić, Werner Streicher, Dominik Heider, Ralf Strasser, Hans Michael Maric |
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URN: | urn:nbn:de:bvb:20-opus-301157 |
Document Type: | Journal article |
Faculties: | Fakultät für Biologie / Rudolf-Virchow-Zentrum |
Language: | English |
Parent Title (English): | Communications Biology |
Year of Completion: | 2022 |
Volume: | 5 |
Article Number: | 1070 |
Source: | Communications Biology (2022) 5:1070. doi:10.1038/s42003-022-03997-3 |
DOI: | https://doi.org/10.1038/s42003-022-03997-3 |
Dewey Decimal Classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
Tag: | combinatorial libraries; kinetics; peptides; screening; thermodynamics |
Release Date: | 2023/04/25 |
Open-Access-Publikationsfonds / Förderzeitraum 2022 | |
Licence (German): | CC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International |