Interrogating the Essential Bacterial Cell Division Protein FtsQ with Fragments Using Target Immobilized NMR Screening (TINS)
Abstract
:1. Introduction
2. Results
2.1. Site-Specific Immobilization of FtsQp
2.2. Fragment Screening Using Target Immobilized NMR Screening (TINS)
2.3. Hit Validation
3. Discussion
4. Materials and Methods
4.1. Expression and Purification of FtsQp
4.2. Immobilization of FtsQp for TINS
4.3. FtsQp Activity Assay
4.4. Target Immobilized NMR screening
4.5. Production of 15N-Labeled FtsQp
4.6. Biosensor Analysis
4.7. Hit Validation by 1H,15N-HSQC NMR
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
Abbreviations
PPI | protein-protein interaction |
NMR | nuclear magnetic resonance |
TINS | target immobilized NMR screening |
SPR | surface plasmon resonance |
HSQC | heteronuclear single quantum coherence |
CSP | chemical shift perturbation |
FBDD | fragment-based drug discovery |
TPSA | total polar surface area |
HTS | high throughput screening |
T/R | target spectrum/reference spectrum ratio |
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Glas, M.; AB, E.; Hollander, J.; Siegal, G.; Luirink, J.; de Esch, I. Interrogating the Essential Bacterial Cell Division Protein FtsQ with Fragments Using Target Immobilized NMR Screening (TINS). Int. J. Mol. Sci. 2019, 20, 3684. https://doi.org/10.3390/ijms20153684
Glas M, AB E, Hollander J, Siegal G, Luirink J, de Esch I. Interrogating the Essential Bacterial Cell Division Protein FtsQ with Fragments Using Target Immobilized NMR Screening (TINS). International Journal of Molecular Sciences. 2019; 20(15):3684. https://doi.org/10.3390/ijms20153684
Chicago/Turabian StyleGlas, Marjolein, Eiso AB, Johan Hollander, Gregg Siegal, Joen Luirink, and Iwan de Esch. 2019. "Interrogating the Essential Bacterial Cell Division Protein FtsQ with Fragments Using Target Immobilized NMR Screening (TINS)" International Journal of Molecular Sciences 20, no. 15: 3684. https://doi.org/10.3390/ijms20153684