Mechanism-Based Strategy for Optimizing HaloTag Protein Labeling

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F22%3A00077616" target="_blank" >RIV/00159816:_____/22:00077616 - isvavai.cz</a>
Alternative codes found
RIV/00216224:14310/22:00126373
Result on the web
<a href="https://pubs.acs.org/doi/10.1021/jacsau.2c00002" target="_blank" >https://pubs.acs.org/doi/10.1021/jacsau.2c00002</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1021/jacsau.2c00002" target="_blank" >10.1021/jacsau.2c00002</a>

Result language
angličtina
Original language name
Mechanism-Based Strategy for Optimizing HaloTag Protein Labeling
Original language description
HaloTag labeling technology has introduced unrivaled potential in protein chemistry and molecular and cellular biology. A wide variety of ligands have been developed to meet the specific needs of diverse applications, but only a single protein tag, DhaAHT, is routinely used for their incorporation. Following a systematic kinetic and computational analysis of different reporters, a tetramethyirhodamine- and three 4-stilbazolium-based fluorescent ligands, we showed that the mechanism of incorporating different ligands depends both on the binding step and the efficiency of the chemical reaction. By studying the different haloalkane dehalogenases DhaA, LinB, and DmmA, we found that the architecture of the access tunnels is critical for the kinetics of both steps and the ligand specificity. We showed that highly efficient labeling with specific ligands is achievable with natural dehalogenases. We propose a simple protocol for selecting the optimal protein tag for a specific Iigand from the wide pool of available enzymes with diverse access tunnel architectures. The application of this protocol eliminates the need for expensive and laborious protein engineering.
Czech name
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Czech description
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Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10400 - Chemical sciences

Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year
2022
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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