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Quantitative Profiling of Dual Phosphorylation of Fus3 MAP Kinase in Saccharomyces cerevisiae
Hur, Jae-Young,Kang, Gum-Yong,Choi, Min-Yeon,Jung, Jin Woo,Kim, Kwang-Pyo,Park, Sang-Hyun Korean Society for Molecular Biology 2008 Molecules and cells Vol.26 No.1
Mitogen-activated protein kinase (MAPK) signaling is a crucial component of eukaryotic cells; it plays an important role in responses to extracelluar stimuli and in the regulation of various cellular activities. The signaling cascade is evolutionarily conserved in the eukaryotic kingdom from yeast to human. In response to a variety of extracellular signals, MAPK activity is known to be regulated via phosphorylation of a conserved TxY motif at the activation loop in which both threonine and tyrosine residues are phosphorylated by the upstream kinase. However, the mechanism by which both residues are phosphorylated continues to remain elusive. In the budding yeast, Saccharomyces cerevisiae, Fus3 MAPK is involved in the mating signaling pathway. In order to elucidate the functional mechanism of MAPK activation, we quantitatively profiled phosphorylation of the TxY motif in Fus3 using mass spectrometry (MS). We used synthetic heavy stable isotope-labeled phosphopeptides and nonphosphopeptides corresponding to the proteolytic TxY motif of Fus3 and accompanying data-dependent tandem MS to quantitatively monitor dynamic changes in the phosphorylation events of MAPK. Phosphospecific immunoblotting and the MS data suggested that the tyrosine residue is dynamically phosphorylated upon stimulation and that this leads to dual phosphorylation. In contrast, the magnitude of threonine phosphorylation did not change significantly. However, the absence of a threonine residue leads to hyperphosphorylation of the tyrosine residue in the unstimulated condition, suggesting that the threonine residue contributes to the control of signaling noise.
Studies on the Iridoid Glycoside and GABA in the Rehmannia glutinosa Cultivars Root
Sang Hoon Lee(이상훈),Jeong Su Yoon(윤정수),Jae Kwang Kim(김재광),Chun Geon Park(박춘근),Mei Lan Jin(김미란),Mok Hur(허목),Seong Cheol Koo(구성철),Woo Moon Lee(이우문),Jae Ki Chang(장재기),Yeon Bok Kim(김연복) 한국약용작물학회 2017 한국약용작물학회 학술대회논문집 Vol.2017 No.2
Aucubin, Catalpol, and GABA Content in Different Organ of Rehmannia glutinosa Cultivars
Sang Hoon Lee(이상훈),Jeong Su Yoon(윤정수),Jae Kwang Kim(김재광),Chun Geon Park(박춘근),Mok Hur(허목),Seong Cheol Koo(구성철),Mei Lan Jin,Woo Moon Lee(이우문),Jae Ki Chang(장재기),Yeon Bok Kim(김연복) 한국약용작물학회 2017 한국약용작물학회 학술대회논문집 Vol.2017 No.1
Characteristics of Eggshell Powder as Carriers of Probiotics
Woo-Do Lee(이우도),Kai-Min Niu(우개민),Jeong-Min Lim(임정민),Kwon-Jung Yi(이권정),Bong-Joo Lee(이봉주),Kang-Woong Kim(김강웅),Kyoung-Duck Kim(김경덕),Sang-Woo Hur(허상우),Hyon-Sob Han(한현섭),Soo-Ki Kim(김수기) 한국생명과학회 2018 생명과학회지 Vol.28 No.1
계란 가공 부산물인 난각(ES: Eggshell)은 탄산칼슘 함량이 높아 사료에 첨가하여 칼슘원으로 이용되고 있다. 본 연구에서는 ES를 생균제의 부형제인 운반체로서 활용 가능성을 처음으로 시도하였다. L. plantarum을 대두박(SBM: Soybean meal), 난각조각(ESL: Eggshell powder with large particles), 난각미세분말(ESF: Eggshell powder with fine particles), 그리고 이들의 복합운반체인 SBM+ESL과 SBM+ESF에 생균제를 흡착시켜 그 부착상태를 주사전자현미경으로 확인하였다. 이 중 복합운반체인 SBM+ESF는 상온에서 4주 동안 pH 7~8을 유지하면서 L. plantarum의 가장 높은 생존율을 보였다. 본 연구에 사용한 모든 생균제들은 보존기간 동안 4℃에서는 높은 생존율을 보였다. 30℃에서는 유산균수는 크게 감소하였으나, B. licheniformis는 높은 생존율을 유지하였고 B. subtilis, B. amyloliquefaciens와 S. cerevisiae는 2 log10 (CFU/g)정도 감소하였다. 상기 연구결과는 사료의 칼슘원으로 이용되는 난각미세분말(ESF)을 대두박과 혼합하여 사용하면 B. licheniformis를 비롯한 일부 생균제의 생존성을 향상시켜 부형제(운반체)로도 사용할 수 있음을 밝혔다. Eggshell (ES) is a by-product of table eggs with high content of calcium carbonate which can be used as a calcium source in feed. In this study, we have first illuminated the potential application of ES as a novel carrier for probiotics. The carriers used in the study include a SBM (Soybean meal), ESL (Eggshell powder with large particles), ESF (Eggshell powder with fine particles), and the complex carriers (SBM+ESL, SBM+ESF). The structure of carriers absorbed by L. plantarum was confirmed by SEM image. Among these carriers, the complex carrier SBM+ESF showed the highest viability of L. plantarum with pH 7~8 during four weeks storage at room temperature. The SBM+ESF was further tested as a carrier for various probiotic strains at 4℃ or 30℃. All the probiotic strains showed high viability at 4℃ storage. However, a significant reduction of Lactobacillus cells was observed at 30℃ storage. B. lichenifomis maintained high viability whereas B. subtilis, B. amyloliquefaciens, and S. cerevisiae showed the reduction of 2 log10 (CFU/g). These results suggest that if the ESF as a calcium source in feed was mixed with SBM, it can be used as an effective complex carrier for improving the viability of some probiotics including B. licheniformis.