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Int. J. Mol. Sci. 2018, 19(10), 2928; https://doi.org/10.3390/ijms19102928

Mapping the Contact Sites of the Escherichia coli Division-Initiating Proteins FtsZ and ZapA by BAMG Cross-Linking and Site-Directed Mutagenesis

1
Mass Spectrometry of Biomacromolecules, Swammerdam Institute for Life Sciences, Faculty of Science, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands
2
Bacterial Cell Biology and Physiology, Swammerdam Institute for Life Sciences, Faculty of Science, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands
3
Computational Structural Biology, Faculty of Science-Chemistry, University of Utrecht, Padualaan 83584CH Utrecht, The Netherlands
4
Faculty of Medical Technology, Prince of Songkla University, Songkhla 90110, Thailand
*
Authors to whom correspondence should be addressed.
Received: 23 August 2018 / Revised: 13 September 2018 / Accepted: 19 September 2018 / Published: 26 September 2018
(This article belongs to the Special Issue Regulatory Mechanisms of Tubulin-Like Proteins)
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Abstract

Cell division in bacteria is initiated by the polymerization of FtsZ at midcell in a ring-like structure called the Z-ring. ZapA and other proteins assist Z-ring formation and ZapA binds ZapB, which senses the presence of the nucleoids. The FtsZ–ZapA binding interface was analyzed by chemical cross-linking mass spectrometry (CXMS) under in vitro FtsZ-polymerizing conditions in the presence of GTP. Amino acids residue K42 from ZapA was cross-linked to amino acid residues K51 and K66 from FtsZ, close to the interphase between FtsZ molecules in protofilaments. Five different cross-links confirmed the tetrameric structure of ZapA. A number of FtsZ cross-links suggests that its C-terminal domain of 55 residues, thought to be largely disordered, has a limited freedom to move in space. Site-directed mutagenesis of ZapA reveals an interaction site in the globular head of the protein close to K42. Using the information on the cross-links and the mutants that lost the ability to interact with FtsZ, a model of the FtsZ protofilament–ZapA tetramer complex was obtained by information-driven docking with the HADDOCK2.2 webserver. View Full-Text
Keywords: cell division; Z associated protein A (ZapA); Filamenting temperature sensitive Z (FtsZ); quadrupole time of flight mass spectrometer (QTOF); Fourier-Transform Ion Cyclotron Resonance mass spectrometry(FTICR); 1,4-bis(succimidyl)-3-azidomethylglutarate (BAMG) cell division; Z associated protein A (ZapA); Filamenting temperature sensitive Z (FtsZ); quadrupole time of flight mass spectrometer (QTOF); Fourier-Transform Ion Cyclotron Resonance mass spectrometry(FTICR); 1,4-bis(succimidyl)-3-azidomethylglutarate (BAMG)
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Roseboom, W.; Nazir, M.G.; Meiresonne, N.Y.; Mohammadi, T.; Verheul, J.; Buncherd, H.; Bonvin, A.M.J.J.; de Koning, L.J.; de Koster, C.G.; de Jong, L.; den Blaauwen, T. Mapping the Contact Sites of the Escherichia coli Division-Initiating Proteins FtsZ and ZapA by BAMG Cross-Linking and Site-Directed Mutagenesis. Int. J. Mol. Sci. 2018, 19, 2928.

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