A Comprehensive Study of the Interaction between Peptidoglycan Fragments and the Extracellular Domain of Mycobacterium tuberculosis Ser/Thr Kinase PknB

Qianqian Wang, Roberta Marchetti, Sladjana Prisic, Kentaro Ishii, Yohei Arai, Ippei Ohta, Shinsuke Inuki, Susumu Uchiyama, Alba Silipo, Antonio Molinaro, Robert N. Husson, Koichi Fukase, Yukari Fujimoto

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The Mycobacterium tuberculosis Ser/Thr kinase PknB is implicated in the regulation of bacterial cell growth and cell division. The intracellular kinase function of PknB is thought to be triggered by peptidoglycan (PGN) fragments that are recognized by the extracytoplasmic domain of PknB. The PGN in the cell wall of M. tuberculosis has several unusual modifications, including the presence of N-glycolyl groups (in addition to N-acetyl groups) in the muramic acid residues and amidation of d-Glu in the peptide chains. Using synthetic PGN fragments incorporating these diverse PGN structures, we analyzed their binding characters through biolayer interferometry (BLI), NMR spectroscopy, and native mass spectrometry (nMS) techniques. The results of BLI showed that muropeptides containing 1,6-anhydro-MurNAc and longer glycan chains exhibited higher binding potency and that the fourth amino acid of the peptide stem, d-Ala, was crucial for protein recognition. Saturation transfer difference (STD) NMR spectroscopy indicated the major involvement of the stem peptide region in the PASTA-PGN fragment binding. nMS suggested that the binding stoichiometry was 1:1. The data provide the first molecular basis for the specific interaction of PGN with PknB and firmly establish PGNs as the effective ligands of PknB.

Original languageEnglish
Pages (from-to)2094-2098
Number of pages5
JournalChemBioChem
Volume18
Issue number21
DOIs
Publication statusPublished - 2017 Nov 2

Keywords

  • biolayer interferometry
  • biomolecular interactions
  • NMR spectroscopy
  • peptidoglycans
  • proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

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  • Cite this

    Wang, Q., Marchetti, R., Prisic, S., Ishii, K., Arai, Y., Ohta, I., Inuki, S., Uchiyama, S., Silipo, A., Molinaro, A., Husson, R. N., Fukase, K., & Fujimoto, Y. (2017). A Comprehensive Study of the Interaction between Peptidoglycan Fragments and the Extracellular Domain of Mycobacterium tuberculosis Ser/Thr Kinase PknB. ChemBioChem, 18(21), 2094-2098. https://doi.org/10.1002/cbic.201700385