Small Mesh Size Hydrogel for Functional Photocontrol of Encapsulated Enzymes and Small Probe Molecules

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• 作者：Shuhei Murayama ; Fumi Ishizuka ; Kaihei Takagi ; Hirotaka Inoda ; Akira Sano ; Tomofumi Santa ; Masaru Kato
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：February 7, 2012
• 年：2012
• 卷：84
• 期：3
• 页码：1374-1379
• 全文大小：275K
• 年卷期：v.84,no.3(February 7, 2012)
• ISSN：1520-6882

文摘

Previously, we developed the 鈥減rotein activation and release from cage by external light鈥?(PARCEL) method for controlling the function of proteins by encapsulating them in a photodegradable hydrogel and subsequently releasing them by ultraviolet (UV) irradiation of the gel. However, controlling small proteins is difficult because small proteins can leak from the gap (ca. 12.4 nm) of the mesh structure of the hydrogel without irradiation. Here, we developed a photodegradable gel with a smaller mesh size (3.6 nm) and used the new gel to control the function of three small enzymes (trypsin, chymotrypsin, and elastase) and several small nonprotein molecules. The new gel showed reduced leakage of the proteins without irradiation, and tryptic activity increased approximately 78-fold upon irradiation of gel-encapsulated trypsin. The new gel also permitted encapsulation and release of 4鈥?6-diamidino-2-phenylindole (DAPI, molecular weight 277), a small DNA-specific fluorescent probe. After irradiation to the gel-encapsulated DAPI and subsequent addition of DNA, strong fluorescence of the DAPI鈥揇NA complex was observed. Our results indicate that reducing the gel mesh size from 12.4 to 3.6 nm should allow the encapsulation of various proteins and small molecules in an inactive state and their subsequent light-induced release. We expect this method to be useful in preparation of photoactivated biosensors, drug delivery systems, and catalysis.