title: Binding mode of N,N',N",N"'-tetraacetylchitotetraitol to hen egg white lysozyme.
abstract: The binding of N,N',N",N"'-tetraacetylchitotetraitol [(GlcNAc)4-ol] to hen egg white lysozyme was investigated by fluorescence and 1H NMR spectroscopy. From observation of changes in the fluorescence intensity, the association constants of (GlcNAc)4-ol and (GlcNAc)3 were found to be 0.70 x 10(5) and 1.07 x 10(5) M-1, respectively, at pH 5.0 and 30 degrees C. The lack of a substantial difference between the association constants suggests that the binding mode of the (GlcNAc)3 moiety of (GlcNAc)4-ol is basically similar to that of (GlcNAc)3, but that the N-acetylglucosaminitol residue of (GlcNAC)4-ol does not interact significantly with lysozyme. On the other hand, 1H NMR spectroscopy revealed a minor difference in the binding modes of the two saccharides. For most of the 1H signals responding to saccharide binding, such as those of Trp 63 H2, Trp 28 H5, and Ile 98 H gamma 1, the chemical shift changes induced by (GlcNAc)4-ol were almost identical to those induced by (GlcNAc)3. However, the effect of binding on the signals of Asn 59 H alpha and Trp 108 indole N1H, which are located near subsite C, was different for (GlcNAc)4-ol and (GlcNAc)3. Thus it is inferred that the binding mode of the first sugar residue of (GlcNAc)4-ol to subsite C is somewhat different from that of (GlcNAc)3.
authors.affiliation: Laboratory of Biophysical Chemistry, Faculty of Agriculture, Kinki University, Nara, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Fukamizo
authors.forename: T
authors.initials: T
authors.lastname: Ohkawa
authors.forename: Y
authors.initials: Y
authors.lastname: Ikeda
authors.forename: T
authors.initials: T
authors.lastname: Torikata
authors.forename: S
authors.initials: S
authors.lastname: Goto
mesh_terms: D000818:Animals; D002240:Carbohydrate Sequence; D002645:Chickens; D004529:Egg White / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009113:Muramidase / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
chemical_list: C092821:N,N',N'',N'''-tetraacetylchitotetraitol; D009844:Oligosaccharides; D009113:Muramidase