Carlsson J, Drevin H, Axén R
Biochem J. 1978 Sep 1;173(3):723-37. doi: 10.1042/bj1730723.
A heterobifunctional reagent, N-succinimidyl 3-(2-pyridyldithio)propionate, was synthesized. Its N-hydroxysuccinimide ester group reacts with amino groups and the 2-pyridyl disulphide structure reacts with aliphatic thiols. A new thiolation procedure for proteins is based on this reagent. The procedure involves two steps. First, 2-pyridyl disulphide structures are introduced into the protein by the reaction of some of its amino groups with the N-hydroxysuccinimide ester sie of the reagent. The protein-bound 2-pyridyl disulphide structures are then reduced with dithiothreitol. This reaction can be carried out without concomitant reduction of native disulphide bonds. The technique has been used for the introduction of thiol groups de novo into ribonuclease, gamma-globulin, alpha-amylase and horseradish peroxidase. N-Succinimidyl 3-(2-pyridyldithio)propionate can also be used for the preparation of protein-protein conjugates. This application is based on the fact that protein-2-pyridyl disulphide derivatives (formed from the reaction of non-thiol proteins with the reagent) react with thiol-containing proteins (with native thiols or thiolated by, for example, the method described above) via thiol-disulphide exchange to form disulphide-linked protein-protein conjugates. This conjugation technique has been used for the preparation of an alpha-amylase-urease, a ribonuclease-albumin and a peroxidase-rabbit anti-(human transferrin) antibody conjugate. The disulphide bridges between the protein molecules can easily be split by reduction or by thiol-disulphide exchange. Thus conjugation is reversible. This has been demonstrated by scission of the ribonuclease-albumin and the alpha-amylase-urease conjugate into their components with dithiothreitol. N-Succinimidyl 3-(2-pyridyldithio)propionate has been prepared in crystalline form, in which state (if protected against humidity) it is stable on storage at room temperature (23 degrees C).
合成了一种异双功能试剂N - 琥珀酰亚胺基3 -（2 - 吡啶基二硫代）丙酸酯。其N - 羟基琥珀酰亚胺酯基团与氨基反应，2 - 吡啶基二硫结构与脂肪族硫醇反应。基于该试剂建立了一种新的蛋白质硫醇化方法。该方法包括两个步骤。首先，通过试剂的N - 羟基琥珀酰亚胺酯基团与蛋白质的一些氨基反应，将2 - 吡啶基二硫结构引入蛋白质中。然后用二硫苏糖醇还原与蛋白质结合的2 - 吡啶基二硫结构。该反应可以在不伴随天然二硫键还原的情况下进行。该技术已用于将巯基从头引入核糖核酸酶、γ - 球蛋白、α - 淀粉酶和辣根过氧化物酶中。N - 琥珀酰亚胺基3 -（2 - 吡啶基二硫代）丙酸酯还可用于制备蛋白质 - 蛋白质缀合物。该应用基于以下事实：蛋白质 - 2 - 吡啶基二硫衍生物（由非硫醇蛋白质与试剂反应形成）通过硫醇 - 二硫交换与含硫醇的蛋白质（具有天然硫醇或通过例如上述方法硫醇化）反应，形成二硫键连接的蛋白质 - 蛋白质缀合物。这种缀合技术已用于制备α - 淀粉酶 - 脲酶、核糖核酸酶 - 白蛋白和过氧化物酶 - 兔抗（人转铁蛋白）抗体缀合物。蛋白质分子之间的二硫键可以通过还原或硫醇 - 二硫交换轻易断裂。因此缀合是可逆的。用二硫苏糖醇将核糖核酸酶 - 白蛋白和α - 淀粉酶 - 脲酶缀合物裂解成其组分已证明了这一点。N - 琥珀酰亚胺基3 -（2 - 吡啶基二硫代）丙酸酯已制成晶体形式，在这种状态下（如果防潮）在室温（23℃）储存时稳定。
