Biomimicry enhances sequential reactions of tethered glycolytic enzymes, TPI and GAPDHS.
Biomimicry enhances sequential reactions of tethered glycolytic enzymes, TPI and GAPDHS.
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DOI：
10.1371/journal.pone.0061434
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Travis AJ
中科院分区：
文献类型：
--
作者：
Mukai C;Gao L;Bergkvist M;Nelson JL;Hinchman MM;Travis AJ
保持酶的活性拴在固体界面仍然是一个主要的挑战，在开发杂化有机-无机器件。在自然界中，哺乳动物精子已经克服了这一设计挑战，通过具有专门的靶向结构域的糖酵解酶，使它们能够在与细胞骨架元件连接时发挥功能。作为一个步骤，朝着设计一个混合的有机-无机ATP生成系统，我们实施了一个仿生位点特异性固定化策略拴两个糖酵解酶代表不同的功能酶家族：丙糖磷酸异构酶（TPI;异构酶）和甘油醛3-磷酸脱氢酶（GAPDHS;氧化还原酶）。然后，我们评估了这些酶的活性相比，当他们通过经典的羧基胺交联栓。无论固定方法如何，两种酶均显示出相似的表面结合。值得注意的是，这两种酶的比活性显着较高时，使用仿生，位点特异性固定化方法拴系。使用这种仿生方法，我们将两种酶连接到单个表面，并在正向和反向方向上连续展示它们的功能。同样，当使用这种仿生方法偶联酶时，与羧基-胺结合相比，在两个方向上串联的活性显著更高。我们的研究结果表明，仿生，位点特异性固定可以提供重要的功能优势，化学特异性，但非定向连接，一个重要的战略洞察力，越来越多的兴趣，概括整个生物途径的混合有机-无机设备。
Maintaining activity of enzymes tethered to solid interfaces remains a major challenge in developing hybrid organic-inorganic devices. In nature, mammalian spermatozoa have overcome this design challenge by having glycolytic enzymes with specialized targeting domains that enable them to function while tethered to a cytoskeletal element. As a step toward designing a hybrid organic-inorganic ATP-generating system, we implemented a biomimetic site-specific immobilization strategy to tether two glycolytic enzymes representing different functional enzyme families: triose phosphoisomerase (TPI; an isomerase) and glyceraldehyde 3-phosphate dehydrogenase (GAPDHS; an oxidoreductase). We then evaluated the activities of these enzymes in comparison to when they were tethered via classical carboxyl-amine crosslinking. Both enzymes show similar surface binding regardless of immobilization method. Remarkably, specific activities for both enzymes were significantly higher when tethered using the biomimetic, site-specific immobilization approach. Using this biomimetic approach, we tethered both enzymes to a single surface and demonstrated their function in series in both forward and reverse directions. Again, the activities in series were significantly higher in both directions when the enzymes were coupled using this biomimetic approach versus carboxyl-amine binding. Our results suggest that biomimetic, site-specific immobilization can provide important functional advantages over chemically specific, but non-oriented attachment, an important strategic insight given the growing interest in recapitulating entire biological pathways on hybrid organic-inorganic devices.
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影响因子：
41.2
作者：
Luo, TJM;Soong, R;Montemagno, C
通讯作者：
Montemagno, C
DOI：
10.1095/biolreprod2.supplement_2.90
发表时间：
1970-01-01
期刊：
Biol. Reprod.
影响因子：
--
作者：
Fawcett, D. W.
通讯作者：
Fawcett, D. W.
影响因子：
2.9
作者：
Halliwell, CM;Morgan, G;Cass, AEG
通讯作者：
Cass, AEG
影响因子：
4.9
作者：
Du, YZ;Hiratsuka, Y;Kodaka, M
通讯作者：
Kodaka, M
影响因子：
3.3
作者：
Travis, AJ;Foster, JA;Moss, SB
通讯作者：
Moss, SB