The Soluble Periplasmic Domains of Escherichia coli Cell Division Proteins FtsQ/FtsB/FtsL Form a Trimeric Complex with Submicromolar Affinity.

M. Glas; H.B. van den Berg van Saparoea; S.H. Mclaughlin; W. Roseboom; F. Liu; G.M. Koningstein; A. Fish; T. den Blaauwen; A.J. Heck; L. de de Jong; W. Bitter; I.J.P. de Esch; S. Luirink

doi:10.1074/jbc.M115.654756

The Soluble Periplasmic Domains of Escherichia coli Cell Division Proteins FtsQ/FtsB/FtsL Form a Trimeric Complex with Submicromolar Affinity.

M. Glas
, H.B. van den Berg van Saparoea
, S.H. Mclaughlin
, W. Roseboom
, F. Liu
, G.M. Koningstein
, A. Fish
, T. den Blaauwen
, A.J. Heck
, L. de de Jong
, W. Bitter
, I.J.P. de Esch
, S. Luirink

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

Abstract

Cell division in Escherichia coli involves a set of essential proteins that assembles at midcell to form the so-called divisome. The divisome regulates the invagination of the inner membrane, cell wall synthesis, and inward growth of the outer membrane. One of the divisome proteins, FtsQ, plays a central but enigmatic role in cell division. This protein associates with FtsB and FtsL, which, like FtsQ, are bitopic inner membrane proteins with a large periplasmic domain (denoted FtsQ<inf>p</inf>, FtsB<inf>p</inf>, and FtsL<inf>p</inf>) that is indispensable for the function of each protein. Considering the vital nature and accessible location of the FtsQBL complex, it is an attractive target for protein-protein interaction inhibitors intended to block bacterial cell division. In this study, we expressed FtsQ<inf>p</inf>, FtsB<inf>p</inf>, and FtsL<inf>p</inf> individually and in combination. Upon co-expression, FtsQ<inf>p</inf> was co-purified with FtsB<inf>p</inf> and FtsL<inf>p</inf> from E. coli extracts as a stable trimeric complex. FtsB<inf>p</inf> was also shown to interact with FtsQ<inf>p</inf> in the absence of FtsL<inf>p</inf> albeit with lower affinity. Interactions were mapped at the C terminus of the respective domains by site-specific cross-linking. The binding affinity and 1:1:1 stoichiometry of the FtsQ<inf>p</inf>B<inf>p</inf>L<inf>p</inf> complex and the FtsQ<inf>p</inf>B<inf>p</inf> subcomplex were determined in complementary surface plasmon resonance, analytical ultracentrifugation, and native mass spectrometry experiments.

Original language	English
Pages (from-to)	21498-21509
Journal	Journal of Biological Chemistry
Volume	290
Issue number	35
DOIs	https://doi.org/10.1074/jbc.M115.654756
Publication status	Published - 2015

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Glas, M., van den Berg van Saparoea, H. B., Mclaughlin, S. H., Roseboom, W., Liu, F., Koningstein, G. M., Fish, A., den Blaauwen, T., Heck, A. J., de Jong, L. D., Bitter, W., de Esch, I. J. P., & Luirink, S. (2015). The Soluble Periplasmic Domains of Escherichia coli Cell Division Proteins FtsQ/FtsB/FtsL Form a Trimeric Complex with Submicromolar Affinity. Journal of Biological Chemistry, 290(35), 21498-21509. https://doi.org/10.1074/jbc.M115.654756

@article{d8c3796446d741bf9ee3e8eb8c0255c1,

title = "The Soluble Periplasmic Domains of Escherichia coli Cell Division Proteins FtsQ/FtsB/FtsL Form a Trimeric Complex with Submicromolar Affinity.",

abstract = "Cell division in Escherichia coli involves a set of essential proteins that assembles at midcell to form the so-called divisome. The divisome regulates the invagination of the inner membrane, cell wall synthesis, and inward growth of the outer membrane. One of the divisome proteins, FtsQ, plays a central but enigmatic role in cell division. This protein associates with FtsB and FtsL, which, like FtsQ, are bitopic inner membrane proteins with a large periplasmic domain (denoted FtsQp, FtsBp, and FtsLp) that is indispensable for the function of each protein. Considering the vital nature and accessible location of the FtsQBL complex, it is an attractive target for protein-protein interaction inhibitors intended to block bacterial cell division. In this study, we expressed FtsQp, FtsBp, and FtsLp individually and in combination. Upon co-expression, FtsQp was co-purified with FtsBp and FtsLp from E. coli extracts as a stable trimeric complex. FtsBp was also shown to interact with FtsQp in the absence of FtsLp albeit with lower affinity. Interactions were mapped at the C terminus of the respective domains by site-specific cross-linking. The binding affinity and 1:1:1 stoichiometry of the FtsQpBpLp complex and the FtsQpBp subcomplex were determined in complementary surface plasmon resonance, analytical ultracentrifugation, and native mass spectrometry experiments.",

author = "M. Glas and \{van den Berg van Saparoea\}, H.B. and S.H. Mclaughlin and W. Roseboom and F. Liu and G.M. Koningstein and A. Fish and \{den Blaauwen\}, T. and A.J. Heck and \{de Jong\}, \{L. de\} and W. Bitter and \{de Esch\}, I.J.P. and S. Luirink",

year = "2015",

doi = "10.1074/jbc.M115.654756",

language = "English",

volume = "290",

pages = "21498--21509",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

publisher = "American Society for Biochemistry and Molecular Biology Inc.",

number = "35",

}

Glas, M, van den Berg van Saparoea, HB, Mclaughlin, SH, Roseboom, W, Liu, F, Koningstein, GM, Fish, A, den Blaauwen, T, Heck, AJ, de Jong, LD, Bitter, W , de Esch, IJP & Luirink, S 2015, 'The Soluble Periplasmic Domains of Escherichia coli Cell Division Proteins FtsQ/FtsB/FtsL Form a Trimeric Complex with Submicromolar Affinity.', Journal of Biological Chemistry, vol. 290, no. 35, pp. 21498-21509. https://doi.org/10.1074/jbc.M115.654756

TY - JOUR

T1 - The Soluble Periplasmic Domains of Escherichia coli Cell Division Proteins FtsQ/FtsB/FtsL Form a Trimeric Complex with Submicromolar Affinity.

AU - Glas, M.

AU - van den Berg van Saparoea, H.B.

AU - Mclaughlin, S.H.

AU - Roseboom, W.

AU - Liu, F.

AU - Koningstein, G.M.

AU - Fish, A.

AU - den Blaauwen, T.

AU - Heck, A.J.

AU - de Jong, L. de

AU - Bitter, W.

AU - de Esch, I.J.P.

AU - Luirink, S.

PY - 2015

Y1 - 2015

N2 - Cell division in Escherichia coli involves a set of essential proteins that assembles at midcell to form the so-called divisome. The divisome regulates the invagination of the inner membrane, cell wall synthesis, and inward growth of the outer membrane. One of the divisome proteins, FtsQ, plays a central but enigmatic role in cell division. This protein associates with FtsB and FtsL, which, like FtsQ, are bitopic inner membrane proteins with a large periplasmic domain (denoted FtsQp, FtsBp, and FtsLp) that is indispensable for the function of each protein. Considering the vital nature and accessible location of the FtsQBL complex, it is an attractive target for protein-protein interaction inhibitors intended to block bacterial cell division. In this study, we expressed FtsQp, FtsBp, and FtsLp individually and in combination. Upon co-expression, FtsQp was co-purified with FtsBp and FtsLp from E. coli extracts as a stable trimeric complex. FtsBp was also shown to interact with FtsQp in the absence of FtsLp albeit with lower affinity. Interactions were mapped at the C terminus of the respective domains by site-specific cross-linking. The binding affinity and 1:1:1 stoichiometry of the FtsQpBpLp complex and the FtsQpBp subcomplex were determined in complementary surface plasmon resonance, analytical ultracentrifugation, and native mass spectrometry experiments.

AB - Cell division in Escherichia coli involves a set of essential proteins that assembles at midcell to form the so-called divisome. The divisome regulates the invagination of the inner membrane, cell wall synthesis, and inward growth of the outer membrane. One of the divisome proteins, FtsQ, plays a central but enigmatic role in cell division. This protein associates with FtsB and FtsL, which, like FtsQ, are bitopic inner membrane proteins with a large periplasmic domain (denoted FtsQp, FtsBp, and FtsLp) that is indispensable for the function of each protein. Considering the vital nature and accessible location of the FtsQBL complex, it is an attractive target for protein-protein interaction inhibitors intended to block bacterial cell division. In this study, we expressed FtsQp, FtsBp, and FtsLp individually and in combination. Upon co-expression, FtsQp was co-purified with FtsBp and FtsLp from E. coli extracts as a stable trimeric complex. FtsBp was also shown to interact with FtsQp in the absence of FtsLp albeit with lower affinity. Interactions were mapped at the C terminus of the respective domains by site-specific cross-linking. The binding affinity and 1:1:1 stoichiometry of the FtsQpBpLp complex and the FtsQpBp subcomplex were determined in complementary surface plasmon resonance, analytical ultracentrifugation, and native mass spectrometry experiments.

U2 - 10.1074/jbc.M115.654756

DO - 10.1074/jbc.M115.654756

M3 - Article

SN - 0021-9258

VL - 290

SP - 21498

EP - 21509

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 35

ER -

The Soluble Periplasmic Domains of Escherichia coli Cell Division Proteins FtsQ/FtsB/FtsL Form a Trimeric Complex with Submicromolar Affinity.

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