title: A study of D52S hen lysozyme-GlcNAc oligosaccharide complexes by NMR spectroscopy and electrospray mass spectrometry.
abstract: The production of a mutant hen lysozyme is described in which Asp-52, one of the catalytically important residues, is replaced by Ser. The mutant enzyme has very low catalytic activity but NMR studies show that its structure is closely similar to that of the wild-type protein. NMR experiments also show that well defined complexes are formed with GlcNAc4 and GlcNAc6 bound in the active site of the mutant enzyme. These complexes have been examined using electrospray mass spectrometry (ESMS). The most intense peaks arise from the uncomplexed protein indicating that dissociation takes place in the mass spectrometer under the conditions used here. Peaks from minor species corresponding to complexes between the protein and the oligosaccharides are, however, also observed. The possibility that the latter arise from novel covalent enzyme-saccharide complexes is discussed.
authors.affiliation: Inorganic Chemistry Laboratory, University of Oxford, UK.
authors.forename: K J
authors.initials: KJ
authors.lastname: Lumb
authors.forename: R T
authors.initials: RT
authors.lastname: Aplin
authors.forename: S E
authors.initials: SE
authors.lastname: Radford
authors.forename: D B
authors.initials: DB
authors.lastname: Archer
authors.forename: D J
authors.initials: DJ
authors.lastname: Jeenes
authors.forename: N
authors.initials: N
authors.lastname: Lambert
authors.forename: D A
authors.initials: DA
authors.lastname: MacKenzie
authors.forename: C M
authors.initials: CM
authors.lastname: Dobson
authors.forename: G
authors.initials: G
authors.lastname: Lowe
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002645:Chickens; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D009113:Muramidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism
chemical_list: D009844:Oligosaccharides; D009113:Muramidase; D000117:Acetylglucosamine