Interrogating the Essential Bacterial Cell Division Protein FtsQ with Fragments Using Target Immobilized NMR Screening (TINS)

Division of Molecular Microbiology, Amsterdam Institute for Molecules, Medicines and Systems (AIMMS),Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands

Division of Medicinal Chemistry, Amsterdam Institute for Molecules, Medicines and Systems (AIMMS),Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands

Zobio BV, 2333 CH Leiden, The Netherlands

Author to whom correspondence should be addressed.

Int. J. Mol. Sci. 2019, 20(15), 3684; https://doi.org/10.3390/ijms20153684

Received: 21 June 2019 / Revised: 20 July 2019 / Accepted: 22 July 2019 / Published: 27 July 2019

(This article belongs to the Section Molecular Microbiology)

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Abstract

The divisome is a large protein complex that regulates bacterial cell division and therefore represents an attractive target for novel antibacterial drugs. In this study, we report on the ligandability of FtsQ, which is considered a key component of the divisome. For this, the soluble periplasmic domain of Escherichia coli FtsQ was immobilized and used to screen a library of 1501 low molecular weight (< 300 Da), synthetic compounds for those that interact with the protein. A primary screen was performed using target immobilized NMR screening (TINS) and yielded 72 hits. Subsequently, these hits were validated in an orthogonal assay. At first, we aimed to do this using surface plasmon resonance (SPR), but the lack of positive control hampered optimization of the experiment. Alternatively, a two-dimensional heteronuclear single quantum coherence (HSQC) NMR spectrum of FtsQ was obtained and used to validate these hits by chemical shift perturbation (CSP) experiments. This resulted in the identification of three fragments with weak affinity for the periplasmic domain of FtsQ, arguing that the ligandability of FtsQ is low. While this indicates that developing high affinity ligands for FtsQ is far from straightforward, the identified hit fragments can help to further interrogate FtsQ interactions. View Full-Text

Keywords: bacterial cell division; antibacterials; Escherichia coli; fragment screening; divisome; FtsQ; NMR; TINS

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MDPI and ACS Style

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

AMA Style

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 Style

Glas, 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

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Interrogating the Essential Bacterial Cell Division Protein FtsQ with Fragments Using Target Immobilized NMR Screening (TINS)

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