Fine Mutational Analysis of 2B8 and 3H7 Tag Epitopes with Corresponding Specific Monoclonal Antibodies

부성희,김태림,Man-Ho Cho,Kanidta Sangsawang 한국분자세포생물학회 2016 Molecules and cells Vol.39 No.6

Bacteriophytochromes are phytochrome-like light-sensing photoreceptors that use biliverdin as a chromophore. To study the biochemical properties of the Deinococcus radi- odurans bacteriophytochrome (DrBphP) protein, two anti- DrBphP mouse monoclonal antibodies (2B8 and 3H7) were generated. Their specific epitopes were identified in our previous report. We present here fine epitope mapping of these two antibodies by using truncation and substitution of original epitope sequences in order to identify mini- mized epitope peptides. The previously reported original epitope sequences for 2B8 and 3H7 were truncated from both sides. Our analysis showed that the minimal peptide sequence lengths for 2B8 and 3H7 antibodies were nine amino acids (RDPLPFFPP) and six amino acids (PGEIEE), respectively. We further characterized these peptides in order to investigate their reactivity after single deletion and single substitution of the original peptides. We found that single-substituted 2B8 epitope (RDPLPAFPP) and dual- substituted 3H7 epitope (PGEIAD) showed significantly increased reactivity. These two antibodies with high reac- tivity for the short modified peptide sequences are value- ble for developing new peptide tags for protein research.

Purification and Characterization of Chloroplast Fructose-1,6-bisphosphatase from Pea Leaves

부성희,한태룡,Bhoo, Seong-Hee,Hahn, Tae-Ryong 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.4

엽록체 fructose-1,6-bisphosphatase를 완두 잎으로부터 1,300배 정제하였다. 정제된 효소는 거의 순수하였고 SDS polyacrylamide 전기영동하여 얻은 monomer의 분자량은 약 40,000이었다. 효소는 pH 7.5에서 활성이 없었으나 dithiothreitol 혹은 알카리 pH에서 활성화가 되었다. 이 결과는 분리된 fructose-1,6-bisphosphatase가 엽록체로부터 얻어진 것임을 나타낸다. Fructose-1,6-phosphate와 $Mg^{++}$에 대한 효소 포회곡선은 Hill 상수 2.6 및 3.1을 가진 sigmoidal한 모양을 보여주는데 이는 효소가 multimer로 구성된 allosteric 한 것임을 알 수 있다. 최고 활성의 반에 요구되는 기질농도는 $40{\mu}M$로서 이는 시금치 엽록체 효소에 대한 $80{\mu}M$값 (Zimmermann et al., 1976)보다 작았다. Chloroplast fructose-1,6-bisphosphatase was purified 1,300 fold from pea leaves. The purified enzyme appeared homogeneous and the approximate molecular weight of the monomer was 40,000, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme was inactive at pH 7.5, while it was activated by dithiothreitol or alkaline pH, indicating that the purified fructose-1,6-bisphosphatase was originated from chloroplast. The enzyme saturation curves with fructose-1,6-bisphosphate and $Mg^{++}$ show sigmoidal shapes with almost same Hill coefficients (2.6 and 3.1, respectively), suggesting that the enzyme is composed of multimer. The substrate concentration required for half-maximal activity was $40{\mu}M$ which is a comparable value $(80{\mu}M)$ for spinach chloroplast fructose bisphosphatase (Zimmermann et al., 1976).

식물 Proteome의 최근 연구동향

부성희,피봉관,전종성 한국유전체학회 2004 유전체소식 Vol.4 No.1

완두엽록체 fructose - 1 , 6 - bisphosphatase 의 분리정제 및 특성

부성희,한태룡 ( Seong Hee Bhoo,Tae Ryong Hahn ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.4

Chloroplast fructose-1,6-bisphosphatase was purified 1,300 fold from pea leaves. The purified enzyme appeared homogeneous and the approximate molecular weight of the monomer was 40,000, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme was inactive at pH 7.5, while it was activated by dithiothreitol or alkaline pH, indicating that the purified fructose-1,6-bisphosphatase was originated from chloroplast. The enzyme saturation curves with fructose-1,6-bisphosphate and Mg^(++) show sigmoidal shapes with almost same Hill coefficients (2.6 and 3.1, respectively), suggesting that the enzyme is composed of multimer. The substrate concentration required for half-maximal activity was 40 μM which is a comparable value (80μM) for spinach chloroplast fructose bisphosphatase (Zimmermann et al., 1976).

Review : Phytochrome signaling compounds and interacting factors involved in Light signaling of Arabidopsis

부성희,신동호 한국생화학분자생물학회 (구 한국생화학회) 2006 생화학분자생물학회 소식 Vol.26 No.4

Deinococcus radiodurans 박테리오피토크롬 특이 항체들의 생산

부성희,김태림,한태룡 한국응용생명화학회 2010 Journal of Applied Biological Chemistry (J. Appl. Vol.53 No.2

To analyze the surface properties of bacteriophytochrome (BphP), five (2B8, 2C11, 3B2, 3D2, 3H7) anti-BphP monoclonal antibodies were produced by using full-length of BphP of Deinococcus radiodurnas. 2B8 and 2C11 preferentially recognized the epitopes at N-terminal region of BphP, whereas 3B2, 3D2 and 3H7 showed preferential affinities to the epitopes of C-terminal region of BphP.

교사 지식 발달에서의 이론-실천 간의 관계: 폴라니 인식론에서의 의미론적 통합을 중심으로

부성희 ( Sung Hee Boo ) 교육철학회 2008 교육철학연구 Vol.42 No.-

연세대학교 의과대학 의학전문직업성 교육 경험의 반성적 고찰

안신기,부성희,An, Shinki,Bu, Sunghee 연세대학교 의과대학 2012 의학교육논단 Vol.14 No.1

For decades medical educators have continually emphasized medical professionalism, which is reflective response to the challenges of a rapidly changing medical environment. This study aimed to review the experience of implementing medical professionalism education at Yonsei University College of Medicine (YUCM). YUCM introduced a new curriculum in 2004 designed by Curriculum Development Project 2004 (CDP2004), a project that was launched in 2001. CDP2004 reorganized lectures as organ-based integrated lectures, introduced an introductory course for clinical medicine and medical humanities courses for premedical and medical students. Problem-based learning (PBL), elective courses, and self-study sessions in the afternoon were implemented in order to equip students with a self-directed learning attitude as medical professionals. Professors were asked by the CDP2004 curriculum to spend more time on student education and to adopt new teaching methods. Experiences of the CDP2004 curriculum reveals 1) difficulty of motivating professors to be PBL tutors 2) students' dissatisfaction with the medical humanities course (major critique was that the course was impractical and unrealistic), and 3) students' optimistic understanding about their future role as medical professionals in influencing and helping people in spite of their perception of the general medical environment not as promising. To foster professionalism, the following are necessary in our experiences: 1) faculty development of medical humanities and medical professionalism, 2) establishment of an environment throughout the whole institution to support medical professionalism education and to integrate the concept into praxis, 3) emphasis on the fact that medical professionalism education is not contradictory to biophysical medical education.

식물세포의 일주기성 단백질

황희연,부성희 한국응용생명화학회 2010 Journal of Applied Biological Chemistry (J. Appl. Vol.53 No.3

In the past 10 years, a lot of plant circadian rhythm researches have published in molecular biology and biochemistry. We discussed with published molecular studies of circadian clock and rhythmic genes in Arabidopsis, rice and algae. However past this studies are not sufficient to explain the whole rhythmic metabolism. Recently many researchers have concerned post-transcriptional, translational and post-translational modification of rhythmic proteins. From the view point of the high-throughput study, we could suggest the proteomic analysis with 2-DE gel electrophoresis and MS/MS techniques for the identification of modified proteins. 지난 10년간 분자 식물 일주기성에 대해 분자 생물학적, 생화학적인 연구가 많이 진행되었다. 본 연구에서는 식물의 Arabidopsis, rice 그리고 algae에서 지금까지 발표된 연구들을 종합하고 고찰해보려 했다. 그 결과, 아직까지도 주기성 대사의 모든 부분을 설명하기엔 부족한 부분이 많다는 것을 알수 있었다. 최근 주기성 단백질들의 전사후, 번역 그리고 번역후 변형과정에 대해 많은 연구자들이 관심을 갖기 시작했다. 이러한 부분에서 앞으로는 다량의 단백질을 한번에 볼 수 있는 2-DE gelelectrophoresis와 MS/MS 기술이 절실히 요구된다고 할 수 있겠다.

피토크롬(Phytochrome)과 피토크롬 유사 단백 질의 Domain 비교 분석

윤주미,부성희 慶熙大學校 食糧資源開發硏究所 2007 硏究論文集 Vol.26 No.2

애기장대 피토크롬 A(AtPhyA), 남조류의 한 종류인 Scyanobacteria 피토크롬 아그로박테리움(Agrobacterium turnefaciens) 피토크롬1(Agp1) Deinococcus radiodurans의 박테리아 피토크롬(DrBphP)의 특징과 그 아미노산 서열의 상동성을 domain 별로 나누어 조사한 결과 전제적인 아미노산 상동성은 222%에서 32%로 나타났으나(Talbe 1), 피토크롬 종류의 단백질이 가지고 있는 특징적인 GAF domin의 아미노산 서열의 상동성은 36%에서 46%로 나타나(Table 2) 진화상이 GAF doamin은 비교적 잘 보존되어 왔다고 생각 할 수 있다. 다음으로 상동성이 높게 나타난 PHY domain역시 피토크롬의 활성화상태를 유지하는데 있어 중요한 역할을 한다고 생각할 수 있을 것이다. GAF domian을 제외한 다른 세 종류의 domain에서 볼 때, 식물 피토크롬과의 상동성은 박테리아와 남조류에서 유래한 피토크롬의 상동성보다 낮게 나타났다. (Table 2). 피토크롬의 아미노산 서열을 바탕으로 계통도를 그려보면 Figure 2와 같으며, 이 계통도를 통해 피토크롬 간의 진화단계상 거리를 알 수 있다. 즉, 식물 피토크롬이 가장 상위에 있다고 할 수 있으며, 단계별로 Agpl, Cphl, 그리고 DrBphP로써 가장 하위에 있는 DrBphP는 피토크롬 종류의 단백질 중 가장 진화되지 않은 형태로 짐작할 수 있을 것이다. 이러한 원형의 피토크롬 단백질들을 연구함으로써 보다 쉽게 광신호 전달 메카니즘에 대한 궁금증을 풀 수 있을 것이다. 이러한 연구가 이루어짐으로써, 앞으로 식물이나 동물의 광신호 전달 메카니즘을 밝히는 초석이 될 수 있을 것이다. 이로써 얻어질 많은 과학적 발전을 생각해 볼 때, 박테리아 유래의 피토크롬에 관한 연구는 앞으로도 계속 이루어져야 할 것으로 생각된다.