Interrogating the essential bacterial cell division protein FtsQ with fragments using target immobilized NMR screening (TINS)

Marjolein Glas, A. B. Eiso, Johan Hollander, Gregg Siegal, Joen Luirink, Iwan de Esch

Research output: Contribution to JournalArticleAcademicpeer-review

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.

Original languageEnglish
Article number3684
JournalInternational Journal of Molecular Sciences
Volume20
Issue number15
DOIs
Publication statusPublished - 1 Aug 2019

Fingerprint

cell division
Cell Division
affinity
Screening
screening
Cells
Nuclear magnetic resonance
fragments
proteins
Proteins
nuclear magnetic resonance
Surface Plasmon Resonance
Chemical shift
Surface plasmon resonance
low molecular weights
Escherichia
surface plasmon resonance
Escherichia coli
Libraries
chemical equilibrium

Keywords

  • Antibacterials
  • Bacterial cell division
  • Divisome
  • Escherichia coli
  • Fragment screening
  • FtsQ
  • NMR
  • TINS

Cite this

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title = "Interrogating the essential bacterial cell division protein FtsQ with fragments using target immobilized NMR screening (TINS)",
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.",
keywords = "Antibacterials, Bacterial cell division, Divisome, Escherichia coli, Fragment screening, FtsQ, NMR, TINS",
author = "Marjolein Glas and Eiso, {A. B.} and Johan Hollander and Gregg Siegal and Joen Luirink and {de Esch}, Iwan",
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Interrogating the essential bacterial cell division protein FtsQ with fragments using target immobilized NMR screening (TINS). / Glas, Marjolein; Eiso, A. B.; Hollander, Johan; Siegal, Gregg; Luirink, Joen; de Esch, Iwan.

In: International Journal of Molecular Sciences, Vol. 20, No. 15, 3684, 01.08.2019.

Research output: Contribution to JournalArticleAcademicpeer-review

TY - JOUR

T1 - Interrogating the essential bacterial cell division protein FtsQ with fragments using target immobilized NMR screening (TINS)

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AU - Eiso, A. B.

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AU - Luirink, Joen

AU - de Esch, Iwan

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AB - 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.

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