有证据表明，木瓜凝乳蛋白酶A的活性中心与其他一些半胱氨酸蛋白酶的活性中心不同，并且硫醇盐阴离子与2,2'-二吡啶二硫化物反应的布仑斯惕系数（β nuc.）可能会因试剂质子化而降低。
Evidence that the active centre of chymopapain A is different from the active centres of some other cysteine proteinases and that the Brønsted coefficient (beta nuc.) for the reactions of thiolate anions with 2,2'-dipyridyl disulphide may be decreased by reagent protonation.
作者信息
Brocklehurst K, Baines B S, Mushiri M S
出版信息
Biochem J. 1980 Jul 1;189(1):189-29. doi: 10.1042/bj1890189.
Abstract
The active centres of chymopapains A and B (jointly designated EC 3.4.22.6) and papaya (Carica papaya L.) peptidase A were investigated by using 2,2'-dipyridyl disulphide and 5,5'-dithiobis-(2-nitrobenzoic acid) as thiol-specific reactivity probes. Whereas the first active-centre pKa values for chymopapain B and papaya peptidase A are less than 5, is as the case for papain (EC 3.4.22.2) and ficin (EC 3.4.22.3), that for chymopapain A is about 6.8. The reason why the reactions of thiols of pKa approx. 6.5 with 2.2'-dipyridyl disulphide are essentially pH-independent in the pH range around the thiol pKa is delineated. The value of the Brønsted coefficient (beta nuc.) for the reactions of thiolate ions with the 2,2'-dipyridyl disulphide monocation appears to be smaller than its value for the corresponding reactions with the neutral disulphide.
摘要
使用2,2'-二吡啶二硫化物和5,5'-二硫代双（2-硝基苯甲酸）作为硫醇特异性反应探针，对凝乳蛋白酶A和B（联合指定为EC 3.4.22.6）以及木瓜（番木瓜）蛋白酶A的活性中心进行了研究。凝乳蛋白酶B和木瓜蛋白酶A的第一个活性中心pKa值小于5，与木瓜蛋白酶（EC 3.4.22.2）和无花果蛋白酶（EC 3.4.22.3）的情况一样，而凝乳蛋白酶A的该值约为6.8。阐述了pKa约为6.5的硫醇与2,2'-二吡啶二硫化物的反应在硫醇pKa附近的pH范围内基本与pH无关的原因。硫醇负离子与2,2'-二吡啶二硫化物单阳离子反应的布伦斯特系数（βnuc.）值似乎小于其与中性二硫化物相应反应的值。
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Evidence that the active centre of chymopapain A is different from the active centres of some other cysteine proteinases and that the Brønsted coefficient (beta nuc.) for the reactions of thiolate anions with 2,2'-dipyridyl disulphide may be decreased by reagent protonation.有证据表明，木瓜凝乳蛋白酶A的活性中心与其他一些半胱氨酸蛋白酶的活性中心不同，并且硫醇盐阴离子与2,2'-二吡啶二硫化物反应的布仑斯惕系数（β nuc.）可能会因试剂质子化而降低。
Biochem J. 1980 Jul 1;189(1):189-29. doi: 10.1042/bj1890189.
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本文引用的文献
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Chymopapain. IV. The chromatographic fractionation of partially purified chymopapain and the characterization of crystalline chymopapain B.
Biochim Biophys Acta. 1967 Jul 11;139(2):405-17. doi: 10.1016/0005-2744(67)90044-7.
2
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。
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3
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A necessary modification to the preparation of papain from any high-quality latex of Carica papaya and evidence for the structural integrity of the enzyme produced by traditional methods.对从任何优质番木瓜乳胶制备木瓜蛋白酶的必要改进以及传统方法生产的酶结构完整性的证据。
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Biochem J. 1979 Nov 1;183(2):223-31. doi: 10.1042/bj1830223.
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Characterization of the papain active centre by using two-protonic-state electrophiles as reactivity probes. Evidence for nucleophilic reactivity in the un-interrupted cysteine-25-histidine-159 interactive system.使用双质子态亲电试剂作为反应性探针表征木瓜蛋白酶活性中心。在不间断的半胱氨酸-25-组氨酸-159相互作用系统中亲核反应性的证据。
Biochem J. 1978 May 1;171(2):385-401. doi: 10.1042/bj1710385.
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