Replacement of Phe274 with conserved residue Tyr274 for reactive center loop expulsion in antithrombin.
Replacement of Phe274 with conserved residue Tyr274 for reactive center loop expulsion in antithrombin.
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DOI：
10.1177/1076029609360529
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发表时间：
2011-06
期刊：
影响因子：
--
通讯作者：
Duhan U
中科院分区：
文献类型：
--
作者：
Duhan U
天然抗凝血酶的反应中心环部分插入丝氨酸蛋白酶抑制剂主体中。为了测试P14-s2 B相互作用影响抗凝血酶中的环插入的假设，它必须完全暴露于最佳抑制功能。通过将Phe 274突变为Tyr 274，目的是在抗凝血酶中引入P14-s2 B相互作用。通过定点突变和亲和层析获得纯化的重组蛋白。测定抗凝血酶与凝血酶形成SDS稳定复合物的能力、凝血酶抑制的化学计量、凝血酶和fXa抑制的二级速率常数（M− 1 s −1）和肝素解离常数（KD）（色氨酸荧光发射光谱）。观察到野生型和突变体之间的微小但不确定的差异。该结果突出了不同丝氨酸蛋白酶抑制剂中P14-s2 B相互作用的可变效应。提出了实现线圈脱出的替代假设。
The Reactive Center Loop of native antithrombin is partially inserted in the main serpin body. It must be fully exposed for optimal inhibitory function To test the hypothesis that P14- s2B interaction affects loop insertion in antithrombin. By mutating Phe274 to Tyr274 the objective was to introduce P14-s2B interaction in antithrombin. Site directed mutagenesis and affinity chromatography were used to obtain purified recombinant protein. Antithrombin’s ability to form SDS stable complex with thrombin, stoichiometry of thrombin inhibition, second order rate constant for thrombin and fXa inhibition (M−1s−1) and heparin dissociation constant (KD) (tryptophan fluorescence emission spectra) were determined. A marginal, but inconclusive, difference between the wild type and mutant was observed. The result highlights the variable effect of P14-s2B interaction in different serpins. Alternate hypothesis for achieving loop expulsion is proposed.
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影响因子：
2.9
作者：
Huntington, JA;Olson, ST;Gettins, PGW
通讯作者：
Gettins, PGW
影响因子：
4.8
作者：
Izaguirre, G;Olson, ST
通讯作者：
Olson, ST
DOI：
10.1016/0006-291x(83)90550-8
发表时间：
1983-01-01
影响因子：
3.1
作者：
CHOAY, J;PETITOU, M;GATTI, G
通讯作者：
GATTI, G
影响因子：
4.8
作者：
Futamura, A;Gettins, PGW
通讯作者：
Gettins, PGW
DOI：
10.1016/s0006-291x(67)80055-x
发表时间：
1967-01-01
影响因子：
3.1
作者：
SCHECHTER, I;BERGER, A
通讯作者：
BERGER, A