NMR and modeling studies of protein-carbohydrate interactions:: Synthesis, three-dimensional structure, and recognition properties of a minimum hevein domain with binding affinity for chitooligosaccharides

被引:62
作者
Aboitiz, N
Vila-Perelló, M
Groves, P
Asensio, JL
Andreu, D
Cañada, FJ
Jiménez-Barbero, J
机构
[1] CSIC, Ctr Invest Biol, Dept Prot Struct & Funct, Madrid 28040, Spain
[2] Univ Pompeu Fabra, Dept Expt & Hlth Sci, Barcelona 08003, Spain
关键词
carbohydrate binding; chitin; hevein; molecular dynamics; NMR spectroscopy;
D O I
10.1002/cbic.200400025
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
HEV32, a 32-residue, truncated hevein locking eleven C-terminal amino acids, was synthesized by solid-phase methodology and correctly folded with three cysteine bridge pairs. The affinities of HEV32 for small chitin fragments-in the forms of N,N',N"-triacetylchitotriose ((GlcNAc)(3)) (millimolar) and N,N',N",N"',N"",N""'-hexaacetylchitohexaose ((GlcNAc)(6)) (micromolar)-as measured by NMR and fluorescence methods, are comparable with those of native hevein. The HEV32 ligand-binding process is enthalpy driven, while entropy opposes binding. The NMR structure of ligand-bound HEV32 in aqueous solution was determined to be highly similar to the NMR structure of ligand-bound hevein.
引用
收藏
页码:1245 / 1255
页数:11
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