以法呢焦磷酸酯螢光類似物探測十一異戊二烯焦磷酸合成酶的活性中心和反應構形變化

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以法呢焦磷酸酯螢光類似物探測十一異戊二烯焦磷酸合成酶的活性中心和反應構形變化

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

李育瑾 , Masters　　Advisor：陳昭岑　　

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法呢焦磷酸酯螢光類似物；十一異戊二烯焦磷酸合成酶； Farnesyl Pyrophosphate Fluorescent Analogues ； Undecaprenyl Pyrophosphate Synthase ；十一異戊二烯焦磷酸合成酶； Farnesyl Pyrophosphate Fluorescent Analogues ； Undecaprenyl Pyrophosphate Synthase

Abstract │ Reference (57) │ Times Cited (4) │ Altmetrics

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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.

Reference ( 57 ) 〈TOP〉

1. Poulter, C. D.; Rilling, H. C. Biosynthesis of Isoprenoid Compounds (Spurgeon, S.L., ed.) John Wiley & Sons, New York. 1981.
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2. Poulter, C. D. J. Agric. Food Chem. 1974, 22, 167-173.
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3. Sinensky, M. Biochim. Biophys. Acta. 2002, 1484, 93-106.
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7. Robyt, J. Essentials of Carbohydrate Chemistry. Springer-Verlag, New York. 1998.
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8. Casey, P. J. Biochem. Soc. Trans. 1995, 23, 161-166.
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