BIOSYNTHESIS OF THE TRYPANOSOMATID METABOLITE TRYPANOTHIONE - PURIFICATION AND CHARACTERIZATION OF TRYPANOTHIONE SYNTHETASE FROM CRITHIDIA-FASCICULATA

被引：35

作者：

HENDERSON, GB ^{[1
]}

YAMAGUCHI, M ^{[1
]}

NOVOA, L ^{[1
]}

FAIRLAMB, AH ^{[1
]}

CERAMI, A ^{[1
]}

机构：

[1] UNIV LONDON LONDON SCH HYG & TROP MED,DEPT MED PARASITOL,LONDON WC1E 7HT,ENGLAND

来源：

BIOCHEMISTRY | 1990年 / 29卷 / 16期

关键词：

D O I：

10.1021/bi00468a019

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：

Trypanothione synthetase from Crithidia fasciculata has been purified ca. 14 500-fold to homogeneity in an overall yield of 40%. The pure enzyme catalyzed the synthesis of N1- and N8-gluta-thionylspermidine and N1,N8-bis(glutathionyl)spermidine (trypanothione) from ATP/magnesium, glutathione (GSH), and spermidine, N1- and N8-glutathionylspermidines being intermediates of trypanothione synthesis. The enzyme showed a sharp pH optimum of 7.5–7.75 for the synthesis of both mono- and diglutathionylspermidine conjugates. It was highly specific for its physiological substrates ATP/Mg2+, GSH, spermidine, and N1- and N8-glutathionylspermidine with Km values of 400 μM, 914 μM, 1.07 mM, 20 μM, and 7 μM, respectively. Trypanothione synthetase was active in the monomeric form with Mr = 87 000 and absorption maxima λmax = 225 and 280 nm (A280/A260 = 1.85). Trypanothione synthetase is a new member of the ATP-dependent class of ligases which form amide linkage with concomitant production of ADP and orthophosphate. © 1990, American Chemical Society. All rights reserved.

引用

页码：3924 / 3929

页数：6

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