Dual binding sites for translocation catalysis by Escherichia coli glutathionylspermidine synthetase

Chien Hua Pai, Bing Yu Chiang, Tzu Ping Ko, Chia Cheng Chou, Cheong Meng Chong, Fang Jiun Yen, Shoujun Chen, James K. Coward, Andrew H.J. Wang, Chun Hung Lin

研究成果: 雜誌貢獻文章同行評審

45 引文 斯高帕斯(Scopus)

摘要

Most organisms use glutathione to regulate intracellular thiol redox balance and protect against oxidative stress; protozoa, however, utilize trypanothione for this purpose. Trypanothione biosynthesis requires ATP-dependent conjugation of glutathione (GSH) to the two terminal amino groups of spermidine by glutathionylspermidine synthetase (GspS) and trypanothione synthetase (TryS), which are considered as drug targets. GspS catalyzes the penultimate step of the biosynthesis - amide bond formation between spermidine and the glycine carboxylate of GSH. We report herein five crystal structures of Escherichia coli GspS in complex with substrate, product or inhibitor. The C-terminal of GspS belongs to the ATP-grasp superfamily with a similar fold to the human glutathione synthetase. GSH is likely phosphorylated at one of two GSH-binding sites to form an acylphosphate intermediate that then translocates to the other site for subsequent nucleophilic addition of spermidine. We also identify essential amino acids involved in the catalysis. Our results constitute the first structural information on the biochemical features of parasite homologs (including TryS) that underlie their broad specificity for polyamines.

原文英語
頁(從 - 到)5970-5982
頁數13
期刊EMBO Journal
25
發行號24
DOIs
出版狀態已發佈 - 12月 13 2006
對外發佈

ASJC Scopus subject areas

  • 神經科學 (全部)
  • 分子生物學
  • 生物化學、遺傳與分子生物學 (全部)
  • 免疫學與微生物學 (全部)

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