Purification and characterization of trypanothione reductase from Crithidia fasciculata, a newly discovered member of the family of disulfide-containing flavoprotein reductases.
Purification and characterization of trypanothione reductase from Crithidia fasciculata, a newly discovered member of the family of disulfide-containing flavoprotein reductases.
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Crithidia fasciculata 中锥硫酮还原酶的纯化和表征，这是一种新发现的含二硫键黄素蛋白还原酶家族的成员。
DOI：
10.1021/bi00360a007
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发表时间：
1986
期刊：
影响因子：
2.9
通讯作者：
Walsh,CT
中科院分区：
文献类型：
--
作者：
Shames,SL;Fairlamb,AH;Cerami,A;Walsh,CT
材料与方法层析树脂来自以下来源：DEAE-Sephacel和2，5 ′-ADP-琼脂糖来自Pharmacia，羟基磷灰石来自Bio-Rad，Ultrogel AcA-44来自LKB。碘[1 - 14 C]乙酰胺（25 mCi mM-1）来自新英格兰核公司。蛋白质分子量标准品获自Sigma。锥硫酮由Graeme亨德森博士（洛克菲勒大学）制备。合成方法的细节将在别处发表（亨德森等人，1986年）。人红细胞谷胱甘肽还原酶是Heiner Schirmer博士（海德堡大学生物化学研究所）的馈赠。所有其他试剂和化学品均为市售最高等级。束状。在24 ℃下在20-L大玻璃瓶中培养束状crithidiafasciculata（野生型），同时进行剧烈通气.培养基含有0.5%酵母提取物、0.4%细菌用胰蛋白胨、2%蔗糖、0.02%硫酸链霉素、0.0002%氯化血红素和2.5mL L-1三乙醇胺（pH 8.0）。生长培养基补充有以下维生素（以毫克/升给出）：烟酰胺，4;泛酸钙，
Materials and MethodsMaterials. Chromatography resins were from the following sources: DEAE-Sephacel and 2,, 5'-ADP-Sepharose were from Pharmacia, hydroxylapatite was from Bio-Rad, and Ultrogel AcA-44 was from LKB. Iodo [l-14C] acetamide (25 mCi mM” 1) was from New England Nuclear. Protein molecular weight standards were obtained from Sigma. Trypanothione was prepared by Dr. Graeme Henderson (The Rockefeller University). Details of the syntheticprocedure will be published elsewhere (Henderson et al., 1986). Human erythrocyte glutathione reductase was a kind gift of Dr. Heiner Schirmer (Biochemistry Institute, University of Heidelberg). All other reagents and chemicals were of the highest grade commercially available.Growth of C. fasciculata. Crithidia fasciculata (wild type) were grown in 20-L carboys at 24 C with vigorous aeration. The medium contained 0.5% yeast extract, 0.4% Bacto-Tryptone, 2% sucrose, 0.02% streptomycin sulfate, 0.0002% hemin, and 2.5 mL L” 1 triethanolamine (pH 8.0). The growth medium was supplemented with the following vitamins (given as milligrams per liter): nicotinamide, 4; calcium pantothenate,