Shimoboji Tsuyoshi, Larenas Edmund, Fowler Tim, Kulkarni Samarth, Hoffman Allan S, Stayton Patrick S
Department of Bioengineering, University of Washington, Seattle, WA 98195; and Genencor International, Inc., Palo Alto, CA 94304, USA.
Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16592-6. doi: 10.1073/pnas.262427799. Epub 2002 Dec 16.
The ability to photoregulate enzyme activities could provide important new opportunities for development of diagnostic assays, sequential bioprocessing, and lab assays in both traditional and microfluidic formats. We show here that the photoinduced changes in the size and hydration of a "smart" polymer chain coil can be used to regulate substrate access and enzyme activity when conjugated to the enzyme at a specific point just outside the active site. The photoresponsive polymers thus serve jointly as antennae and actuators that reversibly respond to distinct optical signals to switch the polymer-enzyme conjugates on and off, and work when the conjugate is free in solution or when immobilized on magnetic beads.
光调节酶活性的能力可为开发传统和微流控形式的诊断检测、顺序生物处理及实验室检测提供重要的新机遇。我们在此表明,“智能”聚合物链线圈在大小和水合作用方面的光致变化,当在活性位点外的特定点与酶共轭时,可用于调节底物的可及性和酶活性。因此,光响应聚合物同时充当天线和致动器,可逆地响应不同的光信号来开启和关闭聚合物 - 酶共轭物,并且当共轭物在溶液中游离或固定在磁珠上时均能发挥作用。