title: LysM domains from Pteris ryukyuensis chitinase-A: a stability study and characterization of the chitin-binding site.
abstract: The LysM domain probably binds peptidoglycans, but how it does so has yet to be described. For this report, we measured the thermal stabilities of recombinant LysM domains derived from Pteris ryukyuensis chitinase-A (PrChi-A) and monitored their binding to N-acetylglucosamine oligomers ((GlcNAc)n) using differential scanning calorimetry, isothermal titration calorimetry, and NMR spectroscopy. We thereby characterized certain of the domains' functional and structural features. We observed that the domains are very resistant to thermal denaturation and that this resistance depends on the presence of disulfide bonds. We also show that the stoichiometry of (GlcNAc)n/LysM domain binding is 1:1. (GlcNAc)5 titration experiments, monitored by NMR spectroscopy, allowed us to identify the domain residues that are critical for (GlcNAc)5 binding. The binding site is a shallow groove formed by the N-terminal part of helix 1, the loop between strand 1 and helix 1, the C-terminal part of helix 2, and the loop between helix 2 and strand 2. Furthermore, mutagenesis experiments reiterate the critical involvement of Tyr72 in (GlcNAc)n/LysM domain binding. Ours is the first report describing the physical structure of a LysM oligosaccharide-binding site based on experimental data.
authors: Division of Plant Sciences, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan.
authors: Takayuki
authors: T
authors: Ohnuma
authors: Shoko
authors: S
authors: Onaga
authors: Katsuyoshi
authors: K
authors: Murata
authors: Toki
authors: T
authors: Taira
authors: Etsuko
authors: E
authors: Katoh
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry* / Q000378:metabolism; D019943:Amino Acid Substitution; D002688:Chitinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary / Q000502:physiology; D032498:Pteridium / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D004220:Disulfides; D009844:Oligosaccharides; D010940:Plant Proteins; D011994:Recombinant Proteins; D002688:Chitinases; D000117:Acetylglucosamine