Mapping the contact sites of the escherichia coli division-initiating proteins FtsZ and ZapA by BAMG cross-linking and site-directed mutagenesis

Winfried Roseboom; Madhvi G. Nazir; Nils Y. Meiresonne; Tamimount Mohammadi; Jolanda Verheul; Hansuk Buncherd; Alexandre M.J.J. Bonvin; Leo J. de Koning; Chris G. de Koster; Luitzen De Jong; Tanneke Den Blaauwen

doi: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

Winfried Roseboom, Madhvi G. Nazir, Nils Y. Meiresonne, Tamimount Mohammadi, Jolanda Verheul, Hansuk Buncherd, Alexandre M.J.J. Bonvin, Leo J. de Koning, Chris G. de Koster, Luitzen De Jong^*, Tanneke Den Blaauwen

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

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.

Original language	English
Article number	2928
Journal	International Journal of Molecular Sciences
Volume	19
Issue number	10
DOIs	https://doi.org/10.3390/ijms19102928
Publication status	Published - 1 Oct 2018

Funding

Funding: T.d.B. and J.V. were supported by the DIVINOCELL project of the European Commission (FP7-Health-2007-B-223431). NYM was supported by the NWO, ALW open program (822.02.019) H.B. received a grant for a Ph. D. program at the University of Amsterdam from the Higher Education Commission of the Royal Thai Government.

Keywords

1,4-bis(succimidyl)-3-azidomethylglutarate (BAMG)
Cell division
Filamenting temperature sensitive Z (FtsZ)
Fourier-transform ion cyclotron resonance mass spectrometry(FTICR)
Quadrupole time of flight mass spectrometer (QTOF)
Z associated protein A (ZapA)

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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. (2018). Mapping the contact sites of the escherichia coli division-initiating proteins FtsZ and ZapA by BAMG cross-linking and site-directed mutagenesis. International Journal of Molecular Sciences, 19(10), Article 2928. https://doi.org/10.3390/ijms19102928

@article{cbab67ae0d354ec3974c0aececfb2b28,

title = "Mapping the contact sites of the escherichia coli division-initiating proteins FtsZ and ZapA by BAMG cross-linking and site-directed mutagenesis",

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.",

keywords = "1,4-bis(succimidyl)-3-azidomethylglutarate (BAMG), Cell division, Filamenting temperature sensitive Z (FtsZ), Fourier-transform ion cyclotron resonance mass spectrometry(FTICR), Quadrupole time of flight mass spectrometer (QTOF), Z associated protein A (ZapA)",

author = "Winfried Roseboom and Nazir, {Madhvi G.} and Meiresonne, {Nils Y.} and Tamimount Mohammadi and Jolanda Verheul and Hansuk Buncherd and Bonvin, {Alexandre M.J.J.} and {de Koning}, {Leo J.} and {de Koster}, {Chris G.} and {De Jong}, Luitzen and {Den Blaauwen}, Tanneke",

year = "2018",

month = oct,

day = "1",

doi = "10.3390/ijms19102928",

language = "English",

volume = "19",

journal = "International Journal of Molecular Sciences",

issn = "1661-6596",

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Roseboom, W, Nazir, MG, Meiresonne, NY, Mohammadi, T, Verheul, J, Buncherd, H, Bonvin, AMJJ, de Koning, LJ, de Koster, CG, De Jong, L & Den Blaauwen, T 2018, 'Mapping the contact sites of the escherichia coli division-initiating proteins FtsZ and ZapA by BAMG cross-linking and site-directed mutagenesis', International Journal of Molecular Sciences, vol. 19, no. 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. / Roseboom, Winfried; Nazir, Madhvi G.; Meiresonne, Nils Y. et al.
In: International Journal of Molecular Sciences, Vol. 19, No. 10, 2928, 01.10.2018.

Research output: Contribution to journal › Article › Academic › peer-review

TY - JOUR

T1 - Mapping the contact sites of the escherichia coli division-initiating proteins FtsZ and ZapA by BAMG cross-linking and site-directed mutagenesis

AU - Roseboom, Winfried

AU - Nazir, Madhvi G.

AU - Meiresonne, Nils Y.

AU - Mohammadi, Tamimount

AU - Verheul, Jolanda

AU - Buncherd, Hansuk

AU - Bonvin, Alexandre M.J.J.

AU - de Koning, Leo J.

AU - de Koster, Chris G.

AU - De Jong, Luitzen

AU - Den Blaauwen, Tanneke

PY - 2018/10/1

Y1 - 2018/10/1

N2 - 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.

AB - 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.

KW - 1,4-bis(succimidyl)-3-azidomethylglutarate (BAMG)

KW - Cell division

KW - Filamenting temperature sensitive Z (FtsZ)

KW - Fourier-transform ion cyclotron resonance mass spectrometry(FTICR)

KW - Quadrupole time of flight mass spectrometer (QTOF)

KW - Z associated protein A (ZapA)

UR - http://www.scopus.com/inward/record.url?scp=85054484906&partnerID=8YFLogxK

U2 - 10.3390/ijms19102928

DO - 10.3390/ijms19102928

M3 - Article

AN - SCOPUS:85054484906

SN - 1661-6596

VL - 19

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

IS - 10

M1 - 2928

ER -

Mapping the contact sites of the escherichia coli division-initiating proteins FtsZ and ZapA by BAMG cross-linking and site-directed mutagenesis

Abstract

Funding

Keywords

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