ENZYMATIC SYNTHESIS OF DIADENOSINE TETRAPHOSPHATE AND DIADENOSINE TRIPHOSPHATE WITH A PURIFIED LYSYL-SRNA SYNTHETASE
ENZYMATIC SYNTHESIS OF DIADENOSINE TETRAPHOSPHATE AND DIADENOSINE TRIPHOSPHATE WITH A PURIFIED LYSYL-SRNA SYNTHETASE
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DOI：
10.1016/0006-291x(66)90415-3
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发表时间：
1966-01-01
影响因子：
3.1
通讯作者：
RANDERATH, K
中科院分区：
文献类型：
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作者：
ZAMECNIK, PC;STEPHENSON, ML;RANDERATH, K
在蛋白质合成中赖氨酸氨基酸活化反应的研究中，我们采用旋光色散（ORD）来寻找l-1赖氨酸：sRNA连接酶（AMP）（EC 6.1）构象可能变化的物理证据。1.6）。当赖氨酸活化在恒温室中在卡里60分光偏振计中进行时，在37 ℃下孵育30分钟期间产生科顿效应。这种效应的出现依赖于赖氨酰-sRNA合成酶、赖氨酸、ATP和Mg++的存在，并且通过提高ATP浓度和通过添加焦磷酸酶来增加该效应。孵育后反应混合物的压力透析显示，可透析材料是造成这种ORD效应的原因（图1）。当将可透析材料冻干，溶于水中，并在聚（乙烯亚胺）-纤维素（PEI-纤维素）上进行薄层色谱（TIC）时（Randerath和Randerath，1965，1966），观察到一种未鉴别的新化合物，猝灭短波紫外光。看来ATP在酶促反应过程中被转化为这种化合物。通过制备型TLC分离几微摩尔的新核苷酸用于随后的分析。该化合物已被鉴定为Pi，P4-二（腺苷-5 '）四磷酸。
In a study of the lysine amino acid activation reaction in protein synthesis, we have employed optical rotatory dispersion(ORD) to look for physicrl evidence of a possible change in conformation of l-1ysine: sRNA ligase(AMP),(EC 6.1. 1.6). When lysine activation was carried out in a thermostated cell in the Cary 60 spectropolarimeter, a Cotton effect developed during a 30 minute incubation at 37OC. The appearance of this effect was dependent on the presence of the lysyl-sRNA synthetase, lysine, ATP, and Mg++, and the effect was increased by raising the ATP concentration and by addition of pyrophosphatase. Pressure dialysis of the reaction mixture following incubation revealed that a dialyzable material was responsible for this ORD effect (Fig. 1). When the dialyzable material was lyophilized, taken up in water, and subjected to thin-layer chromatography(TIC) on poly (ethyleneimine)-cellulose(PEI-cellulose)(Randerath and Randerath, 1965, 1966), an unidentified new compound, quenching short-wave ultraviolet light, was observed. It appeared that ATP was being converted into this compound during the enzymatic reaction. A few umoles of the new nucleotide were isolated by preparative TLC for subsequent analysis. The compound has been identified as Pi, P4-di (adenosine-5') tetraphos-