Probing the Active Site and Conformational Change of Undecaprenyl Pyrophosphate Synthase by Farnesyl Pyrophosphate Fluorescent Analogues

碩士 === 國立臺灣大學 === 化學研究所 === 92 === Undecaprenyl pyrophosphate synthase (UPPs) of E. Coli. catalyzes the condensation of eight molecules of isopentyl pyrophosphate (IPP) with farnesyl pyrophosphate (FPP) to generate C55 undecaprenyl pyrophosphate, which serves as a carrier in transporting lipid II ac...

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Bibliographic Details
Main Authors: Yu Chin Li, 李育瑾
Other Authors: Chao-Tsen Chen
Format: Others
Language:zh-TW
Published: 2004
Online Access:http://ndltd.ncl.edu.tw/handle/89253064417225085411
Description
Summary:碩士 === 國立臺灣大學 === 化學研究所 === 92 === Undecaprenyl pyrophosphate synthase (UPPs) of E. Coli. catalyzes the condensation of eight molecules of isopentyl pyrophosphate (IPP) with farnesyl pyrophosphate (FPP) to generate C55 undecaprenyl pyrophosphate, which serves as a carrier in transporting lipid II across the membrane in bacterial cell wall biosythesis. We successively synthesized four FPP fluorescent analogues: 14 (NBD-FPP), 24 (fluorescein-FPP), 34 (MANT-ESTER-FPP) and 44 (MANT-AMIDE-FPP). We obtained spectroscopic characterization of four analogues by steady-state fluorescence spectrophotometer, and we utilized those to observe the ligand interactions with UPPs. And we also got different kinetic parameters of four analogues. 24 has the best competitive inhibition, and when binding with UPPs, the difference of fluorescent intensity can be seen by eyes. 34 can serve as alternative substrate for UPPs, and could be used as drug screening. Analogues 14, 24, 34, 44 could be used to study different FPP-binding proteins and served as high throughput probes.