通过苯并呋咱与木瓜蛋白酶、无花果蛋白酶、菠萝蛋白酶及低分子量硫醇的反应来评估其作为硫醇基团发色氧化剂的性能。
Evaluation of benzofuroxan as a chromophoric oxidizing agent for thiol groups by using its reactions with papain, ficin, bromelain and low-molecular-weight thiols.
作者信息
Shipton M, Stuchbury T, Brocklehurst K
出版信息
Biochem J. 1977 Mar 1;161(3):627-37. doi: 10.1042/bj1610627.
Abstract
- Benzofuroxan (benzofurazan 1-oxide, benzo-2-oxa-1,3-diazole N-oxide) was evaluated as a specific chromophoric oxidizing agent for thiol groups. 2. Aliphatic thiol groups both in low-molecular-weight molecules and in the enzymes papain (EC 3.4.22.2), ficin (EC 3.4.22.3) and bromelain (EC 3.4.22.4) readily reduce benzofuroxan to o-benzoquinone dixime; potential competing reactions of amino groups are negligibly slow. 3. The fate of the thiol depends on its structure: a mechanism is proposed in which the thiol and benzofuroxan form an adduct which, if steric factors permit, reacts with another molecule of thiol to form a disulphide; when the thiol is located in the active site of a thiol proteinase and steric factors preclude enzyme dinner formation, the adduct reacts instead with water or HO- to form a sulphenic acid; attack on the sulphur atom of the adduct by either a sulphur or oxygen nucleophile releases o-benzoquinone dioxine. 4. Benzofuroxan contains n o proton-binding sites with pKa values in the range 3-10 and probably none in the range 0-14; o-benzoquinone dioxine undergoes a one-proton ionization with pKa=6.75.5. o-benzoquinone dioxime absorbs strongly at wavelengths greater than 410nm, where absorption by benzofuroxan, proteins and simple thiol compounds is negligible; 416 nm is an isosbestic point (epsilon 416 = 5110 litre. mol-1-cm-1); epsilon430=3740+[1460/(1+[H+]/Ka)] where pKa=6.75. 6. The possibility of acid-base catalysis of the oxidation by active-centre histidine residues of the thiol proteinases is discussed.
摘要
苯并呋咱（苯并呋喃 -1- 氧化物，苯并 -2- 恶唑 -1,3- 二氮杂环氧化物）被评估为巯基的一种特异性发色氧化剂。
低分子量分子以及木瓜蛋白酶（EC 3.4.22.2）、无花果蛋白酶（EC 3.4.22.3）和菠萝蛋白酶（EC 3.4.22.4）中的脂肪族巯基能轻易地将苯并呋咱还原为邻苯醌二肟；氨基的潜在竞争反应极其缓慢，可忽略不计。
巯基的反应取决于其结构：提出了一种机制，即巯基与苯并呋咱形成加合物，若空间因素允许，该加合物会与另一分子的巯基反应形成二硫化物；当巯基位于巯基蛋白酶的活性位点且空间因素阻止形成酶二聚体时，加合物反而会与水或氢氧根反应形成亚磺酸；硫或氧亲核试剂对加合物硫原子的进攻会释放出邻苯醌二肟。
苯并呋咱在3 - 10范围内没有质子结合位点，在0 - 14范围内可能也没有；邻苯醌二肟发生单质子电离，pKa = 6.75。
邻苯醌二肟在波长大于410nm处有强烈吸收，此时苯并呋咱、蛋白质和简单巯基化合物的吸收可忽略不计；416nm是等吸收点（ε416 = 5110升·摩尔⁻¹·厘米⁻¹）；ε430 = 3740 + [1460 / (1 + [H⁺] / Ka)]，其中pKa = 6.75。
讨论了巯基蛋白酶活性中心组氨酸残基对氧化进行酸碱催化的可能性。
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Evaluation of benzofuroxan as a chromophoric oxidizing agent for thiol groups by using its reactions with papain, ficin, bromelain and low-molecular-weight thiols.通过苯并呋咱与木瓜蛋白酶、无花果蛋白酶、菠萝蛋白酶及低分子量硫醇的反应来评估其作为硫醇基团发色氧化剂的性能。
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引用本文的文献
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