2,2'-二吡啶二硫化物的中性和阳离子形式对硫醇盐阴离子的反应活性。通过三质子态反应探针检测猕猴桃蛋白酶、木瓜蛋白酶和无花果蛋白酶活性中心之间的差异。
Reactivities of neutral and cationic forms of 2,2'-dipyridyl disulphide towards thiolate anions. Detection of differences between the active centres of actinidin, papain and ficin by a three-protonic-state reactivity probe.
作者信息
Brocklehurst K, Stuchbury T, Malthouse J P
出版信息
Biochem J. 1979 Nov 1;183(2):233-8. doi: 10.1042/bj1830233.
Abstract
The second-order rate constants (k) for the reactions of 2,2'-dipyridyl disulphide (pKa2,45) with 2-mercaptoethanol (pKa9.6) and with benzimidazol-2-ylmethanethiol (pKa values 5.6 and 8.3) were determined at 25 degrees C at I 0.1 by stopped-flow spectral analysis over a wide range of pH. These were used to calculate the pH-independent second-order rate constants (k) for the reactions of neutral 2,2'-dipyridyl disulphide and of its monocation with the 2-mercaptoethanol thiolate anion (associated pKa9.6) and with the benzimidazol-2-ylmethanethiol zwitterion (associated pKa5.6). For both thiolate ions, the rate-enhancement factor (kmonocation/kneutral disulphide) is about 1.5x10(3). The dependence on pH in acidic media of k for the reaction of 2,2'-dipyridyl disulphide with actinidin, the thiol proteinase from Actinidia chinensis, was shown to differ from the forms of pH-dependence observed for the analogous reactions with papain (EC 3.4.22.2) and ficin (3.4.22.3). The reactivity of the 2,2'-dipyridyl disulphide dication and its apparent sensitivity to the presence and location of a positive charge in the attacking thiol are discussed.
摘要
通过停流光谱分析，在25℃、离子强度I = 0.1的条件下，于较宽的pH范围内测定了2,2'-二吡啶二硫化物（pKa2.45）与2-巯基乙醇（pKa9.6）以及与苯并咪唑-2-基甲硫醇（pKa值为5.6和8.3）反应的二级速率常数（k）。这些数据用于计算中性2,2'-二吡啶二硫化物及其单阳离子与2-巯基乙醇硫醇阴离子（相关pKa9.6）以及与苯并咪唑-2-基甲硫醇两性离子（相关pKa5.6）反应的与pH无关的二级速率常数（k）。对于这两种硫醇阴离子，速率增强因子（k单阳离子/k中性二硫化物）约为1.5×10³。结果表明，2,2'-二吡啶二硫化物与中华猕猴桃的硫醇蛋白酶猕猴桃蛋白酶反应的k在酸性介质中对pH的依赖性，与观察到的其与木瓜蛋白酶（EC 3.4.22.2）和无花果蛋白酶（3.4.22.3）类似反应的pH依赖性形式不同。本文讨论了2,2'-二吡啶二硫化物二价阳离子的反应活性及其对进攻硫醇中正电荷的存在和位置的明显敏感性。
相似文献
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Reactivities of neutral and cationic forms of 2,2'-dipyridyl disulphide towards thiolate anions. Detection of differences between the active centres of actinidin, papain and ficin by a three-protonic-state reactivity probe.2,2'-二吡啶二硫化物的中性和阳离子形式对硫醇盐阴离子的反应活性。通过三质子态反应探针检测猕猴桃蛋白酶、木瓜蛋白酶和无花果蛋白酶活性中心之间的差异。
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本文引用的文献
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A novel reactivity of papain and a convenient active site titration in the presence of other thiols.木瓜蛋白酶的一种新反应活性以及在其他硫醇存在下简便的活性位点滴定法。
FEBS Lett. 1970 Jul 29;9(2):113-116. doi: 10.1016/0014-5793(70)80327-1.
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Determination of sulfhydryl groups with 2,2'- or 4,4'-dithiodipyridine.用2,2'-或4,4'-二硫代二吡啶测定巯基
Arch Biochem Biophys. 1967 Mar;119(1):41-9. doi: 10.1016/0003-9861(67)90426-2.
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Anionic proteinase from Actinidia chinensis. Preparation and properties of the crystalline enzyme.中华猕猴桃阴离子蛋白酶。结晶酶的制备及性质
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Reactivities of the various protonic states in the reactions of papain and of L-cysteine with 2,2'- and with 4,4'- dipyridyl disulphide: evidence for nucleophilic reactivity in the un-ionized thiol group of the cysteine-25 residue of papain occasioned by its interaction with the histidine-159-asparagine-175 hydrogen-bonded system.木瓜蛋白酶和L-半胱氨酸与2,2'-二吡啶二硫化物和4,4'-二吡啶二硫化物反应中各种质子态的反应活性：木瓜蛋白酶中半胱氨酸-25残基的未电离硫醇基团因其与组氨酸-159-天冬酰胺-175氢键系统相互作用而产生亲核反应活性的证据。
Biochem J. 1972 Jun;128(2):471-4. doi: 10.1042/bj1280471.
5
The actinidin-catalysed hydrolysis of N-a-benzyloxycarbonyl-L-lysine p-nitrophenyl ester. pH dependence and mechanism.
Eur J Biochem. 1973 Jul 16;36(2):575-82. doi: 10.1111/j.1432-1033.1973.tb02947.x.
6
Reactions of papain and of low-molecular-weight thiols with some aromatic disulphides. 2,2'-Dipyridyl disulphide as a convenient active-site titrant for papain even in the presence of other thiols.木瓜蛋白酶及低分子量硫醇与某些芳香族二硫化物的反应。2,2'-二吡啶基二硫化物作为木瓜蛋白酶便捷的活性位点滴定剂，即使在存在其他硫醇的情况下也适用。
Biochem J. 1973 May;133(1):67-80. doi: 10.1042/bj1330067.
7
Kinetic studies on the thiol protease from Actinidia chinensis.中华猕猴桃硫醇蛋白酶的动力学研究。
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8
The unreliability of estimates of group dissociation constants.基团解离常数估计值的不可靠性。
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9
Structure of actinidin: details of the polypeptide chain conformation and active site from an electron density map at 2-8 A resolution.
J Mol Biol. 1977 Sep 25;115(3):263-77. doi: 10.1016/0022-2836(77)90154-1.
10
The equilibrium assumption is valid for the kinetic treatment of most time-dependent protein-modification reactions.平衡假设对于大多数随时间变化的蛋白质修饰反应的动力学处理是有效的。
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