Design and Synthesis of Sugar‐Responsive Semiartificial Myoglobin Triggered by Modulation of Apoprotein–Cofactor Interactions
Design and Synthesis of Sugar‐Responsive Semiartificial Myoglobin Triggered by Modulation of Apoprotein–Cofactor Interactions
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脱辅基蛋白-辅因子相互作用调节引发的糖响应性半人工肌红蛋白的设计与合成
DOI：
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
S. Shinkai
中科院分区：
文献类型：
--
作者：
I. Hamachi;Y. Tajiri;T. Nagase;S. Shinkai
采用辅因子重组法将苯基硼酸基团作为糖识别位点引入天然肌红蛋白。分光光度pH滴定表明糖诱导的pKa位移的H2O协调的血红素中心的半合成肌红蛋白轴承苯基硼酸（met-Mb（PhBOH）2）。通过圆二色性（CD）和顺磁1H NMR光谱，证明了与苯硼酸位点结合的糖诱导血红素裂隙重排以加强血红素辅因子-脱辅基蛋白相互作用。糖的结合诱导的结构变化随后增强Mb（PhBOH）2的双氧存储活性。这种糖诱导的结构和功能的变化没有发生其他修饰的Mb，没有糖识别单位。有趣的是，具有苯基硼酸单元的随机修饰的Mb没有显示出任何糖响应。在Mb（PhBOH）2中，来自糖结合事件的信息被有效地传递到活性中心，使得活性在糖结合时被有效地改变。总之，分子识别单元作为非天然功能分子的活性位点特异性掺入可以为刺激响应性半合成蛋白质的设计提供一种新的策略。
Phenylboronic acid groups as sugar recognition sites were successfully introduced into native myoglobin by a cofactor-reconstitution method. Spectrophotometric pH titration demonstrated the sugar-induced pKa shift of the H2O coordinated to the heme center of the semisynthetic myoglobin bearing phenyl-boronic acids (met-Mb(PhBOH)2). By means of circular dichroism (CD) and paramagnetic 1H NMR spectroscopies, it was proven that sugars that were bound to phenylboronic acid sites induced the rearrangement of the heme crevice to reinforce the heme cofactor-apoprotein interactions. The structural changes that were induced by the binding of sugars subsequently enhanced the dioxygen storage activity of Mb(PhBOH)2. Such sugar-induced structural and functional changes did not occur for other modified Mbs that had no sugar-recognition units. Interestingly, a randomly modified Mb with phenylboronic acid units did not show any sugar response. In Mb(PhBOH)2, the information from the sugar-binding event was efficiently transmitted to the active center, so that the activity was efficiently altered upon sugar binding. In conclusion, the active site specific incorporation of molecular recognition units as nonnatural functional molecules can provide a novel strategy for the design of stimuli-responsive semisynthetic proteins.
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DOI：
10.1016/0006-291x(81)91500-x
发表时间：
1981
影响因子：
3.1
作者：
LaMar,GN;Kong,SB;Smith,KM;Langry,KC
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DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
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作者：
Vyas,K;Rajarathnam,K;Yu,LP;Emerson,SD;LaMar,GN;Krishnamoorthi,R;Mizukami,H
通讯作者：
Mizukami,H
影响因子：
2.9
作者：
L. Vitello;J. Erman;Mark A Miller;Jimin Wang;J. Kraut
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影响因子：
56.9
作者：
JUDICE, JK;GAMBLE, TR;SCHULTZ, PG
通讯作者：
SCHULTZ, PG
影响因子：
2.9
作者：
ERMAN, JE;VITELLO, LB;KRAUT, J
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KRAUT, J