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用亚铁氰化物对漆树漆酶和2型铜缺失酶进行滴定。铜位点的相互关系。

Titrations with ferrocyanide of japanese-lacquer-tree (Rhus vernicifera) laccase and of the type 2 copper-depleted enzyme. Interrelation of the copper sites.

作者信息

Morpurgo L, Graziani M T, Desideri A, Rotilio G

出版信息

Biochem J. 1980 May 1;187(2):367-70. doi: 10.1042/bj1870367.

Abstract
  1. Redox titrations are reported of the metal centres in Japanese-lacquer-tree (Rhus vernicifera) laccase with ferrocyanide. 2. The redox potential of Type 1 Cu was found to increase with ferrocyanide concentration up to a limiting value similar to that for the Type 1 Cu in Type 2 Cu-depleted enzyme (which is independent of ferrocyanide concentration). 3. The redox potential of the two-electron acceptor (Type 3 Cu) is also independent of ferrocyanide concentration in Type 2 Cu-depleted enzyme and lower than values reported for the native enzyme. 4. The two-electron acceptor is present in the oxidized state in the Type 2 Cu-depleted enzyme, though the latter lacks the 330 nm absorption band. 5. The redox potential of Type 2 Cu also depends on ferrocyanide concentration, at least in the presence of azide. 6. The redox potentials are affected by freezing the solutions and/or addition of azide, the latter binding to Type 2 Cu with affinity dependent on the redox state of the two-electron acceptor.
摘要
  1. 报道了用亚铁氰化物对漆树漆酶中的金属中心进行氧化还原滴定。2. 发现1型铜的氧化还原电位随亚铁氰化物浓度增加而升高,直至达到一个极限值,该极限值与2型铜缺失酶中1型铜的极限值相似(该极限值与亚铁氰化物浓度无关)。3. 在2型铜缺失酶中,双电子受体(3型铜)的氧化还原电位也与亚铁氰化物浓度无关,且低于天然酶报道的值。4. 尽管2型铜缺失酶缺乏330nm吸收带,但双电子受体在该酶中以氧化态存在。5. 2型铜的氧化还原电位也取决于亚铁氰化物浓度,至少在有叠氮化物存在时如此。6. 氧化还原电位受溶液冷冻和/或叠氮化物添加的影响,叠氮化物与2型铜结合,其亲和力取决于双电子受体的氧化还原状态。

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本文引用的文献

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Purification and physico-chemical properties of laccase.漆酶的纯化及其物理化学性质
Biochim Biophys Acta. 1958 Oct;30(1):44-52. doi: 10.1016/0006-3002(58)90239-7.
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Electron transfer reactions of copper proteins.铜蛋白的电子转移反应。
Annu Rev Biophys Bioeng. 1976;5:363-96. doi: 10.1146/annurev.bb.05.060176.002051.

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