Solution structure of the chitin-binding domain 1 (ChBD1) of a hyperthermophilic chitinase from Pyrococcus furiosus.

Solution structure of the chitin-binding domain 1 (ChBD1) of a hyperthermophilic chitinase from Pyrococcus furiosus.
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来自激烈火球菌的超嗜热几丁质酶的几丁质结合域 1 (ChBD1) 的溶液结构。

DOI:
10.1093/jb/mvt104
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发表时间:
2006
影响因子:
2.7
通讯作者:
K. Uegaki
K. Uegaki
中科院分区:
生物学4区
文献类型:
--
作者:
S. Mine;Tsutomu Nakamura;Takaaki Sato;T. Ikegami;K. Uegaki

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来自激烈火球菌的几丁质酶是一种能有效水解α和β结晶几丁质的超嗜热糖苷酶。该几丁质酶具有独特的结构特征;它包含两个催化结构域(AD 1和AD 2)和两个几丁质结合结构域(ChBD 1和ChBD 2)。我们已经确定了结构的ChBD 1,这显着提高了催化域的活性,通过核磁共振光谱。ChBD 1的整体结构具有由三条反平行β链组成的紧凑和球形结构,与归类为碳水化合物结合模块(CBM)家族5的其他蛋白质的结构相似。诱变实验表明,三个溶剂暴露的芳香族残基(Tyr 112,Trp 113和Tyr 123)作为几丁质结合位点。第一次证明了Tyr 123或相应的芳香族残基参与其他CBM。这一结果表明,结合模式可能不同于CBM家族5中的其他几丁质结合结构域。此外,ChBD 1和ChBD 2的结合亲和力是相当不同的,这表明这两种ChBD在有效增加AD 1和AD 2的活性方面各自发挥不同的作用。
A chitinase, from Pyrococcus furiosus, is a hyperthermophilic glycosidase that effectively hydrolyses both α and β crystalline chitin. This chitinase has unique structural features; it contains two catalytic domains (AD1 and AD2) and two chitin-binding domains (ChBD1 and ChBD2). We have determined the structure of ChBD1, which significantly enhances the activity of the catalytic domains, by nuclear magnetic resonance spectroscopy. The overall structure of ChBD1 had a compact and globular architecture consisting of three anti-parallel β-strands, similar to those of other proteins classified into carbohydrate-binding module (CBM) family 5. A mutagenesis experiment suggested three solvent-exposed aromatic residues (Tyr112, Trp113 and Tyr123) as the chitin-binding sites. The involvement of Tyr123 or the corresponding aromatic residues in other CBMs, has been demonstrated for the first time. This result indicates that the binding mode may be different from those of other chitin-binding domains in CBM family 5. In addition, the binding affinities of ChBD1 and ChBD2 were quite different, suggesting that the two ChBDs each play a different role in efficiently increasing the activities of AD1 and AD2.
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