Kinetic and paramagnetic NMR investigations of the inhibition of Streptomyces antibioticus tyrosinase

被引：48

作者：

Bubacco, L ^{[1
]}

Vijgenboom, E ^{[1
]}

Gobin, C ^{[1
]}

Tepper, AWJW ^{[1
]}

Salgado, J ^{[1
]}

Canters, GW ^{[1
]}

机构：

[1] Leiden Univ, Inst Chem, Gorlaeus Labs, NL-2300 RA Leiden, Netherlands

来源：

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC | 2000年 / 8卷 / 1-3期

关键词：

tyrosinase; inhibition; paramagnetism; NMR; dinuclear; copper; antiferromagnetic coupling;

D O I：

10.1016/S1381-1177(99)00064-8

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：

A scaled-up isolation and purification procedure is described for tyrosinase from Streptomyces antibioticus. Kinetic studies of the enzyme catalysed conversion of L-3,4-dihydroxyphenylalanine (L-DOPA) into DOPAchrome show that kojic acid, L-mimosine, p-toluic acid and benzoic acid exhibit competitive inhibition with inhibition constants of 3.4, 30, 1.9 X 10(2) and 8.0 X 10(2) mu M, respectively. Paramagnetic NMR techniques appear well suited to study the binding of inhibitors to the active site. From the variation of the NMR shifts with temperature a value of -2J = 156 +/- 6 cm(-1) is derived for the exchange coupling between the unpaired spins on the two Cu(II) ions in the active site of the met-tyrosinase/kojic acid complex. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：27 / 35

页数：9

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