@article{c0dce0efba2d49d3983f42d2986c584b,
title = "A Comprehensive Study of the Interaction between Peptidoglycan Fragments and the Extracellular Domain of Mycobacterium tuberculosis Ser/Thr Kinase PknB",
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.",
keywords = "NMR spectroscopy, biolayer interferometry, biomolecular interactions, peptidoglycans, proteins",
author = "Qianqian Wang and Roberta Marchetti and Sladjana Prisic and Kentaro Ishii and Yohei Arai and Ippei Ohta and Shinsuke Inuki and Susumu Uchiyama and Alba Silipo and Antonio Molinaro and Husson, {Robert N.} and Koichi Fukase and Yukari Fujimoto",
note = "Funding Information: This work was supported in part by grants-in-aid for scientific research from Japan Society for the Promotion of Science (JSPS) (Y.F.; JP26102732, JP26282211, and JP16H01162), NEXT Program from JSPS (Y.F.; LR025), grant from Mizutani Foundation for Gly-coscience (A.M., A.S., and R.M. in 2014; Y.F. in 2016), U.S. National Institutes of Health grant (R.N.H.; R21 AI062275 and R01 AI099204), the Joint Studies Program (2015–2016) in the Okazaki BIO-NEXT project of the Okazaki Institute for Integrative Bioscience (K.I., S.Y, and Y.F.), and also the Osaka University Scholarship for Overseas Research Activities 2014 (Q.W.). We would like to thank Prof. Yoshiaki Furukawa (Keio University) for help with protein expression and also acknowledge Dr. Masaki Sato (Pall Corp.) and Takeshi Seguchi (Primetech Corp.) for assistance in BLI analysis. Publisher Copyright: {\textcopyright} 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim",
year = "2017",
month = nov,
day = "2",
doi = "10.1002/cbic.201700385",
language = "English",
volume = "18",
pages = "2094--2098",
journal = "ChemBioChem",
issn = "1439-4227",
publisher = "Wiley-VCH Verlag",
number = "21",
}