期刊
CHEMBIOCHEM
卷 18, 期 21, 页码 2094-2098出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201700385
关键词
biolayer interferometry; biomolecular interactions; NMR spectroscopy; peptidoglycans; proteins
资金
- Japan Society for the Promotion of Science (JSPS) [JP26102732, JP26282211, JP16H01162]
- NEXT Program from JSPS [LR025]
- Mizutani Foundation for Glycoscience
- U.S. National Institutes of Health grant [R21 AI062275, R01 AI099204]
- Joint Studies Program in the Okazaki BIO-NEXT project of the Okazaki Institute for Integrative Bioscience
- Osaka University Scholarship for Overseas Research Activities
- Grants-in-Aid for Scientific Research [16H00770, 16H01885, 17K19201, 16H01162, 15H05836] Funding Source: KAKEN
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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据