Structural and Functional Study of Glutathione S-transferase Complexed with various substrate

• Main Authors: Yi-Hung Yeh, 葉義弘
• Other Authors: Chuan-Deng Hsiao
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/78906421970823508463

碩士 === 中國文化大學 === 生物科技研究所 === 87 === The crystal structure of chicken mu class glutathione S-transferase M1-1 complexed with a substrate, EPNP(1,2-epoxy-3-(p-nitrophenoxy)propane, has been determined and refined to a resolution 2.8 A. There are two copies of the dimeric enzyme in the asymmetry unit. Each monomer is built from two domain. A bound substrate, EPNP, is primarily associated with one of these domains via a network hydrogen bond and salt bridages. In particular, the nitrogen and oxygen atom of the EPNP forms hydrogen bond with Gln165 and Arg107.In addition, crystals of glutathione S-transferase complexed with the product, PGE2(prostanglandin E2), has been obtained by the vapor diffusion method. The crystal of GST/PGE2 complex has unit cell dimensionsin a = 125.50 A, b = 135.65 A, c = 127.17 A, and diffract to 3.4 A resolution. An analysis of reflection data indicates that the crystal system is orthorhombic, space group C2221. In this thesis, we also report the prelimilary data of mutant crystal of glutathione S-transferase mutant (Arg107 R Leu), has been obtained by the vapor diffusion method. The crystal of GST mutant has unit cell dimensions a = 72.219 A, b = 96.661 A, c = 70.16 A, and diffract to 2.4 A resolution. An analysis of reflection data indicates that the crystal system is orthorhombic, space group P21212.We are trying to improve the crystal quality now.