Developing chemical probe for activity profile of serine hydrolases

• Main Authors: Yu Hsieh, 謝瑀
• Other Authors: Chao-Hsiung Lin
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/28946423398059175235

碩士 === 國立陽明大學 === 生命科學暨基因體科學研究所 === 97 === Serine hydrolases represent one of the largest and diverse families of enzymes comprising numerous proteases, lipases, esterases, and amidases. Serine hydrolases play important roles and also regulate numerous important functions in many organisms. Disorder of such serine hydrolases may cause different kinds of human diseases. To understand the functions of diverse serine hydrolases, it is necessary to develop a method to systematically detect or enrich serine hydrolases and thus the following studies of their biological significances. Fluorophosphonate/fluorophosphates (FP) is a well-known enzyme inhibitors of serine hydrolases. It has been demonstrated to covalently react with the catalytic serine residue of serine hydrolases and thus irreversibly inhibit the enzymatic activity. We collaborated with other laboratory to prepare an FP analogue with the function of biotin and polyethylene glycol (FP-peg-biotin) as a chemical probe to identify serine hydrolases. The following verification used SDS-PAGE, Western blot, and NeutrAvidin beads to separate, detect and enrich serine hydrolases. In summary, we anticipate this probe will help us to develop a high efficient and specific toll for studies of diverse serine hydrolases.