Quantitative Assay for the Binding of Jun-Fos Dimer and Activator Protein-1 Site

Yang, Chul Hak,Jun, Gyo,Lee, Dug Keun,Lee, Sang Kyou,Park, Se Yeon,Hahm, Eun Ryeong 생화학분자생물학회 1998 BMB Reports Vol.32 No.6

The Jun and Fos families of eukaryotic transcription factors form heterodimers capable of binding to their cognate DNA enhancer elements. We are interested in searching for inhibitors or antagonists of the binding of the Jun-Fos heterodimer to the activator protein-1 (AP-1) site. The basic-region leucine zipper (bZIP) domain of c-Fos was expressed as a fusion protein with glutathione S-transferase, and allowed to form a heterodimer with the bZIP domain of c-Jun. The heterodimer was bound to glutathione-agarose, to which were added radiolabeled AP-1 nucleotides. After thorough washing, the gel-bound radioactivity was counted. The assay is faster than the conventional electrophoretic mobility shift assay because the gel electrophoresis step and the autoradiography step are eliminated. Moreover, the assay is very sensitive, allowing the detection of picomolar quantities of nucleotides, and is not affected by up to 50% dimethylsulfoxide, a solvent for hydrophobic inhibitors. Curcumin and dihydroguaiaretic acid, recently known inhibitors of Jun-Fos-DNA complex formation, were applied to this Jun-GST-fused Fos system and revealed to decrease the dimer-DNA binding.

Quantitative Assay for the Binding of Jun-Fos Dimer and Activator Protein-1 Site

Lee, Sang-Kyou,Park, Se-Yeon,Jun, Gyo,Hahm, Eun-Ryeong,Lee, Dug-Keun,Yang, Chul-Hak Korean Society for Biochemistry and Molecular Biol 1999 Journal of biochemistry and molecular biology Vol.32 No.6

The Jun and Fos families of eukaryotic transcription factors form heterodimers capable of binding to their cognate DNA enhancer elements. We are interested in searching for inhibitors or antagonists of the binding of the Jun-Fos heterodimer to the activator protein-1 (AP-1) site. The basic-region leucine zipper (bZIP) domain of c-Fos was expressed as a fusion protein with glutathione S-transferase, and allowed to form a heterodimer with the bZIP domain of c-Jun. The heterodimer was bound to glutathione-agarose, to which were added radiolabeled AP-1 nucleotides. After thorough washing, the gel-bound radioactivity was counted. The assay is faster than the coventional electrophoretic mobility shift assay because the gel electrophoresis step and the autoradiography step are eliminated. Moreover, the assay is very sensitive, allowing the detection of picomolar quantities of nucleotides, and is not affected by up to 50% dimethylsulfoxide, a solvent for hydrophobic inhibitors. Curcumin and dihydroguaiaretic acid, recently known inhibitors of Jun-Fos-DNA complex formation, were applied to this Jun-GST-fused Fos system and revealed to decrease the dimer-DNA binding.

Protein kinase A-induced phosphorylation at the Thr154 affects stability of DJ-1

Ko, Yeon Uk,Kim, Su-Jeong,Lee, Juhyung,Song, Min-Young,Park, Kang-Sik,Park, Jun Bae,Cho, Hyun-Soo,Oh, Young J. Elsevier 2019 Parkinsonism & related disorders Vol.66 No.-

<P><B>Abstract</B></P> <P><B>Introduction</B></P> <P>Most cases of Parkinson's disease (PD) are sporadic, but genetic variations have been discovered in PD patients. PARK7/DJ-1 is a known cause of early-onset autosomal-recessive PD and is implicated in neuroprotection against oxidative stress. Although several post-translational modifications of DJ-1 have been proposed, phospho-modification of DJ-1 and its functional consequences have been less studied.</P> <P><B>Methods</B></P> <P>Putative phosphorylation sites of DJ-1 were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS analysis). Subsequently, phosphorylation site of DJ-1 was confirmed by <I>in vitro</I> kinase assay and cell-based pull-down assay. Impaired dimer formation of phospho-null mutant was measured using DSS crosslinking assay and immunoprecipitation assay. To evaluate physiological consequences of this event, protein stability of DJ-1 WT and DJ-1 phospho-null mutant were compared using cycloheximide chase assay and ubiquitination assay.</P> <P><B>Results</B></P> <P>Here, we showed that DJ-1 directly bound to the catalytic subunit of protein kinase A (PKAcα). We found that PKAcα is responsible for phosphorylation of DJ-1 at the T154 residue. Interestingly, dimerization of DJ-1 was not detected in a DJ-1 T154A mutant. Furthermore, stability of the DJ-1 T154A mutant was dramatically reduced compared with that of wild-type DJ-1. We found that DJ-1 T154A was prone to degradation by the ubiquitin proteasome system (UPS).</P> <P><B>Conclusion</B></P> <P>We identified a novel phosphorylation site of DJ-1. Furthermore, we determined protein kinase A that is responsible for this posttranslational modification. Finally, we demonstrated physiological consequences of this event focusing on dimerization and protein stability of DJ-1.</P> <P><B>Highlights</B></P> <P> <UL> <LI> PKAcα phosphorylates PARK7/DJ1 at the Thr154 residue. </LI> <LI> Disruption of Thr154 phosphorylation is linked to a lower stability. </LI> </UL> </P>

지리산국립공원 심원계곡 생태경관 복원공사지역 식생 변화

정태준 ( Tae-jun Jung ),김연경 ( Yeon-Gyeong Kim ),김영진 ( Young-jin Kim ),정명희 ( Myung-hee Jung ),박경희 ( Kyoung-hee Park ),신창근 ( Chang-keun Shin ),박선홍 ( Seung-hong Park ),김영선 ( Young-sun Kim ) 한국환경생태학회 2021 한국환경생태학회지 Vol.35 No.3

본 연구는 지리산국립공원 심원계곡 생태경관 복원공사지역의 모니터링 결과를 분석하여 체계적인 복원관리를 도모할 수 있는 기초자료를 확보하는데 그 목적이 있다. 복원사업이 완료된 2017년도에 식생 모니터링을 위한 조사구 12개소와 대조구 4개소를 설치하였고, 2017년과 2020년의 상대우점치, 종다양도지수, 유사도 등의 변화를 분석하였다. 복원사업 시 수목이 식재되었던 조사구는 종다양도지수가 0∼1.4552로 나타났고, 대조구와의 유사도지수에서는 1개 조사구가 1.32%로 분석된 것을 제외하고 모두 0%로 나타났다. 조사구의 종다양도지수 및 유사도지수가 매우 낮은 것은 복원사업 시 식재된 수목이 고사하거나 멧돼지 피해, 유수에 의한 침식으로 유실되는 등의 영향으로 판단되었다. 반면 복원사업 시 수목이 식재되지 않은 미식재 조사구는 종다양도지수가 0.9538∼2.3222로 나타났고, 유사도지수는 최대 8.33%까지 분석되었다. 국립공원 생태경관 복원공사지역의 복원 공법 개발을 위한 장기 모니터링과, 미식재 조사구의 천이과정을 분석하는 지속적인 연구가 필요하다. This study aims to obtain basic data for systematic restoration by analyzing the monitoring results of the Shimwon Valley Ecological Landscape Restoration Project area in Jirisan National Park. In 2017, when the restoration project was completed, 12 monitoring plots and 4 control plots were installed for vegetation monitoring, and changes in the relative dominance, species diversity index and similarity between 2017 and 2020 were analyzed. The species diversity index of the surveyed areas where trees were planted during the restoration project was 0-1.4552, and the similarity index with the control group was 0% except for one survey area at 1.32%. The very low species diversity index and similarity index in the survey areas were attributed to the loss of trees planted during the restoration project due to death, damage by wild boars, or erosion by running water. On the other hand, the species diversity index was 0.9538-2.3222 in the monitoring plot where no tree was planted, and the similarity index was analyzed to be as high as 8.33%. It is necessary to continue the long-term monitoring for the development of ecological landscape restoration methods in the national park and analysis of the succession in monitoring plots where no trees were planted.

벤치컷으로 굴착되는 터널의 해석에 적합한 하중분배율의 산정

박연준 水原大學校 2012 論文集 Vol.26 No.-

Load distribution factors suitable for the analyses of tunnels excavated by bench cut were determined. To achieve this, 3-D analyses were conducted while taking every construction stage into account. Then 2-D analyses were conducted which yield the same results with the 3-D results. The crown settlement normalized by the ultimate value was compared during the process to overcome the discrepancy caused by difference in dimension. When the length of the bench is very short compared to the diameter of the tunnel, in such cases as in short bench cut or in mini-bench cut, the supporting effect of the face does not disappear even after the bench is completely excavated since the face is still too close to the point of interest. The 4th load distribution factor was defined to stand for the advance of the face after the completion of the excavation cycle. The 4th load distribution factor turned out to be very useful in analyzing the behavior of tunnels excavated by bench cut with various bench lengths.

FSK 변조에 의한 CPM DS-SS 통신시스템의 단일주파수 잡음에 대한 성능평가

박종연,황준원 江原大學校 産業技術硏究所 1997 産業技術硏究 Vol.17 No.-

This paper has analyzed the performance against a single tone jamming for the CPM DS-SS system with a Binary FSK Modulation. The CPM DS-SS system designed with the different phase shift of the same pseude-noise code has the advantage of reducing the jamming and interference effects on multi-users communication environment. Analytical results show that the system preformance improves by increasing parrallel users in he presence of the coherent single-tone, which is the worst interference since the jamming signal has the same frequency as a carrier.

株式配當의 課稅論

朴英俊 건국대학교 행정대학원 1984 建國大學校 行政大學院 硏究論叢 Vol.12 No.-

ImmunoDot법에 의한 전신성 류마티스 질환의 자가항체 검사

박성환,김선준,김성일,민준기,이상헌,홍연식,조철수,김호연 최신의학사 1997 最新醫學 Vol.40 No.7

Glass-ionomer 시멘트 이장재의 5급와동 치은측 변연폐쇄효과

박연준,손호현 全北大學校 齒醫學硏究所 1987 전북치대논문집 Vol.5 No.1

The purpose of this study is to evaluate the gingival margin sealing effect of glass-ionomer lining cements in Class V cavities. Freshly extracted 120 human teeth were divided into 4 groups. V-shaped cavity was prepared on the cervical area of facial surface of each tooth. Except the 1 ㎜ inner area from the gingival margin, exposed dentin surface was lined with Dycal in Group 1, Lining cement in Group 2 and Chembond cement in Group 3. No liner was used in Group 4. Prepared enamel surface in Group 1 and Group 4 was etched. In Group 2 and Group 3, prepared enemel surface and lined cement surface were etched simultaneously. After etching, washing and drying, bonding agent was applied and cured, and composite resin was inserted and cured with light in every cavity of teeth of all Groups. Restored teeth were thermocycled altenatively in 2% methlene blue dye solution of 4℃ and 60℃. After that, with diamomd disk under water spray, they were sectioned faciolingually to be divided equally. Dye penetration on the sectioned surface was observed with stereomicroscope. After statistical analysis, the following results were obtained : 1. There was significant difference in gingival margin sealing effect in lined cavities, except those lined with Chembond cement, compared with unlined cavities (p<0.01). 2. There was no significant difference in gingival margin sealing effect of glass-ionomer lining cements such as Lining cement and Chembond cement compared with that of Dycal (p>0.05).

脇痛의 鍼灸治療穴에 對한 文獻的 考察

朴基榮,李浚龜,金永一,朴泰均,辛泳一,黃載然,李賢,李秉烈 대전대학교 한의학연구소 2002 한의학연구소 논문집 Vol.10 No.2

As mentioned above, I have acquired some valuable results about medical treatment with acupuncture and Moxibustion of " Flank pain" after studying oriental medical books . The results Were like below: 1. Medical treatment with acupunctures of Flank pain belonged to the Urinary Bladder Meridian of Foot Taiyang, the Liver Meridian of Foot Jueyin, the Gall Bladder Meridian of Foot Shaoyang. 2. Medical treatmcnt with acupunctures of Flank pain used to Yang-laung-chan(陽陵泉), Gi-gu(支溝), Gi-mun(期門), Kan-su(肝兪) in turn. 3. Medical treatment with Ear acupunctures of Flank pain used to Dam(膽), Sin-Mun(神門), Gan(肝)in turn 4. Acupuncture point of Flank pain were lowrer limb part. chest and abdominal part, back part in turn. 5. Medical treatment with Moxibustion of Flank pain was the most Jang-mun(章門)