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Mycobacterium tuberculosis Cell Division Protein, FtsE, is an ATPase in Dimeric Form

The Protein Journal volume 34, pages 35–47 (2015)Cite this article

Abstract

FtsE is one of the earliest cell division proteins that assembles along with FtsX at the mid-cell site during cell division in Escherichia coli. Both these proteins are highly conserved across diverse bacterial genera and are predicted to constitute an ABC transporter type complex, in which FtsE is predicted to bind ATP and hydrolyse it, and FtsX is predicted to be an integral membrane protein. We had earlier reported that the MtFtsE of the human pathogen, Mycobacterium tuberculosis, binds ATP and interacts with MtFtsX on the cell membrane of M. tuberculosis and E. coli. In this study, we demonstrate that MtFtsE is an ATPase, the active form of which is a dimer, wherein the participating monomers are held together by non-covalent interactions, with the Cys84 of each monomer present at the dimer interface. Under oxidising environment, the dimer gets stabilised by the formation of Cys84–Cys84 disulphide bond. While the recombinant MtFtsE forms a dimer on the membrane of E. coli, the native MtFtsE seems to be in a different conformation in the M. tuberculosis membrane. Although disulphide bridges were not observed on the cytoplasmic side (reducing environment) of the membrane, the two participating monomers could be isolated as dimers held together by non-covalent interactions. Taken together, these findings show that MtFtsE is an ATPase in the non-covalent dimer form, with the Cys84 of each monomer present in the reduced form at the dimer interface, without participating in the dimerisation or the catalytic activity of the protein.

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Abbreviations

ABC:

ATP binding cassette

ATP:

Adenosine triphosphate

ADP:

Adenosine diphosphate

Fts:

Filamenting temperature sensitive

NBD:

Nucleotide binding domain

TMD:

Trans membrane domain

DTT:

Dithiothreitol

IPTG:

Isopropylthio-β-d-galactoside

LB broth:

Luria–Bertani broth

PBS:

Phosphate buffered saline

PMSF:

Phenyl methyl sulphonyl fluoride

CuP:

Cupric phenanthroline

DTSP:

3,3′-Dithio-bispropionic acid di (N-hydroxysuccinimide ester)

SDS-PAGE:

Sodium dodecyl sulphate-polyacrylamide gel electrophoresis

β-ΜΕ:

β-Mercaptoethanol

MBP:

Maltose binding protein

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Acknowledgments

MAM thanks Prof. Keith M. Derbyshire, Division of Genetics, Wadsworth Center, Albany, New York, USA, for critical comments on the manuscript. This work was carried out using the ICMR research Grant No. 63/72/2001-BMS to PA and the infrastructural facilities provided by the DBT-supported Genomics Initiative on Microbial Pathogens–Structural Genomics Initiative in the Division of Biological Sciences, Indian Institute of Science, the DST-FIST and UGC-CAS at the Department of Microbiology and Cell Biology, Indian Institute of Science. The authors acknowledge the facilities at the erstwhile DBT-funded Bioinformatics Centre at IISc.

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Affiliations

  1. Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore, 560012, India

    Mushtaq Ahmad Mir, Muthu Arumugam, Haryadi S. Rajeswari & Parthasarathi Ajitkumar

  2. Department of Physics and Bioinformatics Centre, Indian Institute of Science, Bangalore, 560012, India

    Sukanta Mondal & Suryanarayanarao Ramakumar

  3. Division of Genetics, New York State Department of Health, Wadsworth Center, Albany, NY, 12201, USA

    Mushtaq Ahmad Mir

  4. CSIR-National Institute for Interdisciplinary Science and Technology, Thiruvananthapuram, 695019, Kerala, India

    Muthu Arumugam

  5. Biological Sciences, Birla Institute of Technology and Science, Pilani K. K. Birla Goa Campus, Zuarinagar, Goa, 403726, India

    Sukanta Mondal

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  1. Mushtaq Ahmad Mir
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  2. Muthu Arumugam
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  5. Suryanarayanarao Ramakumar
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  6. Parthasarathi Ajitkumar
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Correspondence to Parthasarathi Ajitkumar.

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Mir, M.A., Arumugam, M., Mondal, S. et al. Mycobacterium tuberculosis Cell Division Protein, FtsE, is an ATPase in Dimeric Form. Protein J 34, 35–47 (2015). https://doi.org/10.1007/s10930-014-9593-7

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  • DOI: https://doi.org/10.1007/s10930-014-9593-7

Keywords

  • FtsE and FtsX
  • ATPase
  • Cell division protein
  • Mycobacterium tuberculosis