PREPARATION AND PROPERTIES OF MUTANT TOBACCO PEROXIDASE WITH ADDITIONAL TRYPTOPHAN RESIDUES

A.A.Poloznikov; P.A.Savitskii; D.M.Khushpulyan

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PREPARATION AND PROPERTIES OF MUTANT TOBACCO PEROXIDASE WITH ADDITIONAL TRYPTOPHAN RESIDUES

机译：带有其他色氨酸残基的突变型烟草过氧化物酶的制备及性质

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The Gln116Trp and Leu157Trp mutant tobacco peroxidase forms were prepared by the polymerase chain reaction and expressed by cloning into the pET40b vector and cultivating in E.coli cells BL21(DE3) Codon Plus.The level of both forms of tobacco peroxidase apoprotein expression in E.coli was higher than 40% of the total protein.The refolding of the mutated forms was performed using the procedure known for recombinant wild-type tobacco peroxidase.Mutated tobacco peroxidase reactivation yields were 3-9%,and the specific ABTS activity was 2000-3000 unitsag protein.The substrate concentration dependence of the rate of ABTS oxidation with hydrogen peroxide was studied.The reaction was found to be catalyzed by the mutated forms and follow the ping-pong mechanism.The specific activities of Glnll6Trp and Leul57Trp with respect to various substrates were,however,substantially different.The suggestion can be made that tryptophan residues participate in charge transfer between enzymes and substrate active centers.

机译：通过聚合酶链反应制备Gln116Trp和Leu157Trp突变型烟草过氧化物酶，并将其克隆到pET40b载体中并在大肠杆菌BL21（DE3）Codon Plus中培养表达。两种形式的烟草过氧化物酶载脂蛋白在E中的表达水平。大肠杆菌高于总蛋白的40％。使用已知的重组野生型烟草过氧化物酶的方法进行突变形式的重折叠。经突变的烟草过氧化物酶的重新活化率为3-9％，ABTS的比活为2000- 3000单位蛋白质/ nag蛋白质然而，可以认为色氨酸残基参与了酶与底物之间的电荷转移。 e活动中心。

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《Moscow University Chemistry Bulletin》 |2006年第1期|共6页
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A.A.Poloznikov; P.A.Savitskii; D.M.Khushpulyan;
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1. 叶绿体腺三磷酶中色氨酸残基的修饰与性质的研究 [J] . 王国强, 钱月琴植物学报：英文版 . 1993,第012期
2. PREPARATION AND PROPERTIES OF MUTANT TOBACCO PEROXIDASE WITH ADDITIONAL TRYPTOPHAN RESIDUES [J] . A.A.Poloznikov, P.A.Savitskii, D.M.Khushpulyan Moscow University Chemistry Bulletin . 2006,第1期

机译：带有其他色氨酸残基的突变型烟草过氧化物酶的制备及性质
3. The reactivity of functional groups as a probe for investigating the topography of tobacco mosaic virus. The use of mutants with additional lysine residues in the coat protein [J] . Richard N. Perham The biochemical journal . 1973,第1期

机译：官能团的反应性作为研究烟草花叶病毒形貌的探针。外壳蛋白中带有其他赖氨酸残基的突变体的使用
4. Interaction of tobacco mosaic virus protein with synthetic polynucleotides containing a fluorescent label: optical properties of poly (A,εA) and poly (C,εC) copolymers and energy migration from the tryptophan to 1,N6 or 3,N4-ethenocytosine residues in RNP [J] . A.A. Bogdanov, A.A. Kost, A.P. Razjivin, Nucleic acids research . 1978,第11期

机译：烟草叶片病毒蛋白与含有荧光标记的合成多核苷酸的相互作用：聚（a，εaaa）和聚（c，εc）共聚物的光学性质和从色氨酸至1，n6或3，RNP中的N4- ethenocytosine残基
5. PHOTOSYNTHETIC ELECTRON TRANSPORT PROPERTIES OF THE rbcL DELETION MUTANT OF TOBACCO [C] . Yagut Allahverdiyeva, Imre Vass, Pirkko Maenpaa, International Congress of Photosynthesis . 2005

机译：烟草RBCL缺失突变体的光合电子传输性能
6. Part I. Magnetic circular dichroism studies of cytochrome c peroxidase from yeast: Investigations of the active site heme coordination sphere through comparison with horseradish peroxidase and myoglobin. Part II. Examination of the heme active site coordination structure of the cavity mutants of myoglobin (H93G), cytochrome c peroxidase (H175G), and heme oxygenase (H25A) and structural investigation of thiolate-ligated cytochrome c peroxidase in the H17C/D235L double mutant. [D] . Pond, Alycen Easterling. 1999

机译：第一部分。酵母中细胞色素C过氧化物酶的磁性圆二色性研究：通过与辣根过氧化物酶和肌红蛋白的比较，研究了活性部位血红素配位域。第二部分检查肌红蛋白（H93G），细胞色素c过氧化物酶（H175G）和血红素加氧酶（H25A）的空腔突变体的血红素活性位点配位结构，以及H17C / D235L双突变体中硫醇盐连接的细胞色素c过氧化物酶的结构研究。
7. The reactivity of functional groups as a probe for investigating the topography of tobacco mosaic virus. The use of mutants with additional lysine residues in the coat protein [O] . Richard N. Perham 1973

机译：官能团的反应性作为研究烟草花叶病毒形貌的探针。外壳蛋白中带有其他赖氨酸残基的突变体的使用
8. The role of conserved extracellular cysteine residues in vasopressin V2 receptor function and properties of two naturally occurring mutant receptors with additional extracellular cysteine residues [O] . Schülein Ralf, Zühlke Kerstin, Oksche Alexander, 2000

机译：保守的细胞外半胱氨酸残基在血管加压素V2受体功能中的作用以及两个具有额外细胞外半胱氨酸残基的天然突变受体的特性
9. Energetics of Cation Radical Formation at the Proximal Active Site Tryptophan of Cytochrome-c-Peroxidase and Ascorbate Peroxidase [R] . Jensen, G. M. , Bunte, S. W. , Warshel, A. , 1998

机译：细胞色素-c-过氧化物酶和抗坏血酸过氧化物酶的近端活性位点色氨酸阳离子形成的能量学

PREPARATION AND PROPERTIES OF MUTANT TOBACCO PEROXIDASE WITH ADDITIONAL TRYPTOPHAN RESIDUES

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