尽管在酸性和碱性介质中存在差异，但木瓜蛋白酶和无花果蛋白酶在近中性介质中的催化位点具有高度相似性的证据。木瓜蛋白酶和无花果蛋白酶与氯乙酸反应的动力学。
Evidence for a close similarity in the catalytic sites of papain and ficin in near-neutral media despite differences in acidic and alkaline media. Kinetics of the reactions of papain and ficin with chloroacetate.
作者信息
Brocklehurst K, Mushiri S M, Patel G, Willenbrock F
出版信息
Biochem J. 1982 Jan 1;201(1):101-4. doi: 10.1042/bj2010101.
Abstract
- The pH-dependences of the second-order rate constants (k) for the alkylation by chloroacetate of the active-centre thiol groups of papain (EC 3.4.22.2) and ficin (EC 3.4.22.3) were determined over a wide range of pH at 25 degrees C at I 0.1. 2. The main feature of both pH-k profiles is a striking rate maximum at pH6 (characterizing parameters in both cases pKI approx. 3.5, pKII approx. 8.4 and pH-independent rate constant approximately kXH 2.5-3.0 M-1 . s-1). 3. The profile for the ficin reaction contains a plateau at high pH, with approximately kX 0.10 M-1 . s-1; if an analogous plateau exists in the papain reaction, approximately kX ix much lower, less than 0.02 M-1 . s-1. 4. Both enzymes appear to contain closely similar thiolate-imidazolium interactive systems at pH6, but differences in their behaviour in more-acidic media and in alkaline media suggest differences in interaction with the postulated carboxylate component of the putative catalytic triad.
摘要
- 在25℃、离子强度I = 0.1的条件下，于较宽的pH范围内测定了氯乙酸对木瓜蛋白酶（EC 3.4.22.2）和无花果蛋白酶（EC 3.4.22.3）活性中心巯基进行烷基化反应的二级速率常数（k）的pH依赖性。2. 两种pH - k曲线的主要特征是在pH6时出现显著的速率最大值（两种情况下的特征参数：pKI约为3.5，pKII约为8.4，与pH无关的速率常数约为kXH 2.5 - 3.0 M-1·s-1）。3. 无花果蛋白酶反应的曲线在高pH时包含一个平台，kX约为0.10 M-1·s-1；如果木瓜蛋白酶反应中存在类似的平台，kX则要低得多，小于0.02 M-1·s-1。4. 两种酶在pH6时似乎都含有非常相似的硫醇盐 - 咪唑𬭩相互作用系统，但它们在酸性更强的介质和碱性介质中的行为差异表明，与假定催化三联体中推测的羧酸盐成分的相互作用存在差异。
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1
Evidence for a close similarity in the catalytic sites of papain and ficin in near-neutral media despite differences in acidic and alkaline media. Kinetics of the reactions of papain and ficin with chloroacetate.尽管在酸性和碱性介质中存在差异，但木瓜蛋白酶和无花果蛋白酶在近中性介质中的催化位点具有高度相似性的证据。木瓜蛋白酶和无花果蛋白酶与氯乙酸反应的动力学。
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2
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引用本文的文献
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THE CHEMICAL REACTIVITY OF THE THIOL GROUP IN THE ACTIVE CENTRE OF FICIN.无花果蛋白酶活性中心硫醇基团的化学反应性
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Evidence for a two-state transition in papain that may have no close analogue in ficin. Differences in the disposition of cationic sites and hydrophobic binding areas in the active centres of papain and ficin.木瓜蛋白酶中可能在无花果蛋白酶中没有类似情况的双态转变的证据。木瓜蛋白酶和无花果蛋白酶活性中心中阳离子位点和疏水结合区域分布的差异。
Biochem J. 1980 Dec 1;191(3):707-18. doi: 10.1042/bj1910707.
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Stereospecific alkylation with asymmetric reagents.使用不对称试剂进行立体专一性烷基化反应。
Eur J Biochem. 1968 Jan;3(3):267-75. doi: 10.1111/j.1432-1033.1968.tb19526.x.
4
Reaction of the sulfhydryl group of papain with chloroacetic acid.木瓜蛋白酶的巯基与氯乙酸的反应。
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5
Reaction of chloroacetamide with the sulfhydryl group of papain.氯乙酰胺与木瓜蛋白酶巯基的反应。
J Biol Chem. 1969 Oct 10;244(19):5087-94.
6
Reaction of papain with -bromo- -(5-imidazolyl)propionic acid.木瓜蛋白酶与β-溴-β-(5-咪唑基)丙酸的反应。
Biochemistry. 1972 Jan 18;11(2):164-9. doi: 10.1021/bi00752a005.
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On the mode of activation of the catalytically essential sulfhydryl group of papain.关于木瓜蛋白酶催化必需巯基的激活模式。
Eur J Biochem. 1973 Feb 15;33(1):104-9. doi: 10.1111/j.1432-1033.1973.tb02660.x.
8
Evidence for multiple reactive forms of papain.木瓜蛋白酶多种反应形式的证据。
Eur J Biochem. 1978 Aug 1;88(2):513-21. doi: 10.1111/j.1432-1033.1978.tb12477.x.
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Mechanism of Action of papain with a specific anilide substrate.木瓜蛋白酶与特定酰苯胺底物的作用机制。
Biochemistry. 1979 May 29;18(11):2355-63. doi: 10.1021/bi00578a034.
10
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。
Biochem J. 1979 Feb 1;177(2):541-8. doi: 10.1042/bj1770541.
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