https://scholars.lib.ntu.edu.tw/handle/123456789/146051
標題: | Pirin Regulates Pyruvate Catabolism by Interacting with the Pyruvate Dehydrogenase E1 Subunit and Modulating Pyruvate Dehydrogenase Activity | 作者: | SOO, PO-CHI HORNG, YU-TZE LAI, MENG-JIUN WEI, JIUN-RUNG HSIEH, SHANG-CHEN CHANG, YUNG-LIN TSAI, YU-HUAN LAI, HSIN-CHIH |
關鍵字: | CELL-ENVELOPE ARCHITECTURE;ESCHERICHIA-COLI;MULTIENZYME COMPLEX;SERRATIA-MARCESCENS;SWARMING MOTILITY;NUCLEAR- PROTEIN | 公開日期: | 2007 | 卷: | v.189 | 期: | n.1 | 起(迄)頁: | 109-118 | 來源出版物: | JOURNAL OF BACTERIOLOGY | 摘要: | The protein pirin, which is involved in a variety of biological processes , is conserved from prokaryotic microorganisms, fungi, and plants to mammals. It acts as a transcriptional cofactor or an apoptosis-related protein in mammals and is involved in seed germination and seedling development in plants. In prokaryotes, while pirin is stress induced in cyanobacteria and may act as a quercetinase in Escherichia coli, the functions of pirin orthologs remain mostly uncharacterized. We show that the Serratia marcescens pirin (pirin(Sm)) gene encodes an ortholog of pirin protein . Protein pull-down and bacterial two-hybrid assays followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrospray ionization-tandem mass spectrometry analyses showed the pyruvate dehydrogenase (PDH ) E1 subunit as a component interacting with the pirin(Sm), gene. Functional analyses showed that both PDH El subunit activity and PDH enzyme complex activity are inhibited by the pirin(Sm) gene in S. marcescens CH-1. The S. marcescens CH-1 pirin(Sm) gene was subsequently mutated by insertion- deletion homologous recombination. Accordingly, the PDH Ell and PDH enzyme complex activities and cellular ATP concentration increased up to 250%, 140%, and 220%, respectively, in the S. marcescens CH-1 pirin(Sm) mutant. Concomitantly, the cellular NADH/ NAD(+) ratio increased in the pirin(Sm) mutant, indicating increased tricarboxylic acid (TCA) cycle activity. Our results show that the pirin( Sm) gene plays a regulatory role in the process of pyruvate catabolism to acetyl coenzyme A through interaction with the PDH El subunit and inhibiting PDH enzyme complex activity in S. marcescens CH-1, and they suggest that pirin(Sm) is an important protein involved in determining the direction of pyruvate metabolism towards either the TCA cycle or the fermentation pathways. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/89797 |
顯示於: | 醫學檢驗暨生物技術學系 |
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