Achrobacter Protease I ( API )의 기질특이성의 전환

임성일,최청 ( Seong Il Lim,Cheong Choi ) 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.40 No.3

Assuming that Asp225 is the substrate specificity determinant of Achromobacter protease I (API) which is lysine-specific serine protease, the 225th residue was substituted for other amino acids with a hope that the substrate specificity of a mutant API is altered. Furthermore, to maturate proform of mutant API autocatalytically, Lys(-1) was also replaced by Met, Asp, or Glu. However, all the mutants were not expressed, or accumulated as inactive precursor proteins. This result implicats that Asp225 plays a critical rol in restricted substrate specificity as a lysylendopeptidase but the substrate specificity of API is not determined only by the nature of residue 225.

Achrobacter Protease I (API)의 기질특이성의 전환

임성일,최청,Lim, Seong-Il,Choi, Cheong 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.43 No.2

Lysine 특이적 serine protease인 Achromobacter protease I(API)의 기질특이성을 결정하는 아미노산 잔기가 255위치에 존재하는 가정하에 이 잔가에 변이를 도입하여 기질특이성의 변화 유무를 조사하였다. 그리고 pro-API이 자기촉매적으로 소화될 수 있도록 Lys(-1) 또는 다른 아미노산 잔기로 치환하였다. 그러나 예상과는 달리 제작된 모든 변이체는 발현되지 않았거나 불활성전구체로서 발현되었다. 이 결과로부터 225위치의 잔기는 API의 maturation과정에 있어 Iysylendopeptidase활성에 중요한 역할을 담당하고 있으나 APl의 기질특이성은 225위치 잔기의 특성에 한정되지 않을 가능성이 시사되었다. Assuming that Asp225 is the substrate specificity determinant of Achromobacter pretense I (APl) which is lysine-specific serine protease, the 225th residue was substituted for other amino acids with a hope that the substrate specificity of a mutant API is altered. Furthermore, to maturate preform of mutant API autocatalytically, Lys(-1) was also replaced by Met, Asp, or Glu. However, all the mutants were not expressed, or accumulated as inactive precursor proteins. This result implicats that Asp225 plays a critical rol in restricted substrate specificity as a lysylendopeptidase but the substrate specificity of API is not determined only by the nature of residue 225.

Achromobacter Protease 1 ( API )의 기질특이성 결정기의 동정과 변이체 (D225E)의 Ornithine 기질에 대한 촉매활성

임성일,권오진,최청 ( Seong Il Lim,Oh Jin Kwon,Cheong Choi ) 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.40 No.3

The structural basis of lysine specificity of Achromobacter protease I (API) was investigated by means of site-directed mutagenesis. The precursor protein in which G1u190, one of the two candidates for determining lysine specificity, was substituted by glutamine, aspartic acid or leucine was processed autocatalytically to attain full protease activity with lysine specificity. The substitution of the other candidate, Asp225, for asparagine or leucine produced no mature active forms of pro-API. The precursor protein of the mutant D225E slowly matured autocatalytically. The lysylendopeptidase activity of the mature D225E was 0.25% of that of native API, and this reduced activity is mainly due to a decrease in the affinity of the enzyme for lysine. These results suggest that Asp225 plays a critical rol in restricted substrate specificity as a lysylendopeptidase. However, D225E exhibited no measurable activity for synthetic ornithine substrate. Since the hydroxyl group of Ser194 in this mutant retained essentially the same reactivity to DFP or PMSF as that in native API, it can be noted that a methylene unit longer side chain of residue 225 is not compensated by a methylene unit shorter side chain at subsite P1 in the bound substrate.

Achromobacter Protease I (API)의 기질특이성 결정기의 동정과 변이체[D225E]의 Ornithine 기질에 대한 촉매활성

임성일,권오진,최청,Lim, Seong-Il,Kwon, Oh-Jin,Choi, Cheong 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.43 No.2

부위특이적 변이제작을 통하여 Achromobacter protease I (API)의 lysin에 대한특이성의 구조적 근거를 조사하였다. Lysine 특이성을 결정하는 결정기로서 예상되는 잔기중의 하나인 Glu190을 glutamine, aspartic acid, leucine으로 치환하였을 경우, 전구체단백질은 Iysine 특이성을 가진 완전한 활성효소로서 발현되어 자기촉매적으로 processing되었다. 다른 하나의 잔기인 Asp225을 asparagine, leucine으로 치환한 변이체에서는 불활성 전구체인 pro-API으로서 발현되었으나, 변이체 (D225E)의 경우는 서서히 자기촉매적으로 maturation 되었다. 성숙체인 변이체 (D225E)의 Iysylendopeptidase 활성은 wild-type APl의 0.25%이였으며, 이 활성의 감소는 기질 Iysine과 효소와의 친화력 감소가 주 원인이었다. 이러한 결과로부터 Asp225가 Iysylendopeptidase의 엄격한 기질특이성에 결정적인 역할을 담당한다는 것이 시사되었다. 한편, 변이체 (D225E)는 ornithine 기질에 대해서 효소활성 이 검출되지 않았다. 이 변이체의 촉매잔기인 Ser194가 native API과 마찬가지로 DFP, PMSF와 근본적으로 동일한 반응성이 있는 것으로 나타나 225번 위치의 methylene 1 unit 더 긴 측사는 subsite P1의 methylene 1 unit 짧은 측사에 의해 보상되지 않는다는 것을 알 수 있었다. The structural basis of Iysine specificity of Achromobacter protease I (API) was investigated by means of site-directed mutagenesis. The precursor protein in which Glu190, one of the two candidates for determining Iysine specificity, was substituted by glutamine, aspartic acid or leucine was processed autocatalytically to attaln full pretense activity with lysine specificity. The substitution of the other candidate, Asp225, for asparagine or leucine produced no mature active forms of pro-API. The precursor protein of the mutant D225E slowly matured autocatalytically. The lysylendopeptidase activity of the mature D225E was 0.25% of that of native API, and this reduced activity is mainly due to a decrease in the affinity of the enzyme for lysine. These results suggest that Asp225 plays a critical rol in restricted substrate specificity as a lysylendopeptidase. However, D225E exhibited no measurable activity for synthetic ornithine substrate. Since the hydroxyl group of Ser194 in this mutant retained essentially the same reactivity to DFP or PMSF as that in native API, it can be noted that a methylene unit longer side chain of residue 225 is not compensated by a methylene unit shorter side chain at subsite P1 in the bound substrate.

생물학적 유동층 공법을 이용한 양어장 순환수 내의 질산성질소의 제거

임성일,김정숙,강임석,이병헌 ( Sung Ill Lim,Jeong Soo Kim,Lim Seok Kang,Byung Hun Lee ) 한국물환경학회 1996 학술연구발표회 논문초록집 Vol.1996 No.-

F-E9 : 유류사고 발생시 막여과 시설 운영방안

임성일 ( Sung Il Lim ),이영주 ( Young Joo Lee ),이경혁 ( Kyung Hyuk Lee ),임재림 ( Jae Lim Lim ) 한국물환경학회(구 한국수질보전학회) 2013 한국물환경학회·대한상하수도학회 공동 춘계학술발표회 Vol.2013 No.-

연계선로가 없는 배전선로에서 BESS기반 정전복구를 위한 토폴로지 재구성 방안

임성일(Seongil Lim) 대한전기학회 2017 전기학회논문지 Vol.66 No.10

Outage areas can be transferred to adjacent feeders to restore power supply in case of the fault on the distribution system. Feeders in the small island or mountain area may not have backup feeders due to the low density of load. In this weakly meshed open loop system, BESS can be used as a backup feeder to improve reliability of power supply. This paper proposes a new network reconfiguration method for BESS based service restoration. Fuzzy decision making technique is adopted to deal with fuzziness of service restoration planning rules. Case studies using KEPCO real distribution system have been performed to verify feasibility of the proposed method.

지방자치단체 예산회계제도의 혁신방향

임성일(Sung Il Lim) 한국지방행정연구원 2000 地方行政硏究 Vol.14 No.1

N/A

지방교육재정제도의 개선방안: 지방재정과 지방교육재정간의 관계 재정립

임성일 ( Sung Il Lim ),손희준 ( Hee June Sohn ) 한국지방행정연구원 2011 地方行政硏究 Vol.25 No.3

The purpose of the article is to find ways for the improvement of local education financing system. The current local education financing system experiences low independent status, heavily depending upon the grants(fiscal transfers) from the central government. Under the situation, it is very difficult for local education authorities to respond to the rapidly increasing fiscal needs of local education. The worst problem is the two major equalization grant systems(Local Shared Tax and Local Shared Tax for Education) for local finance operated by the different line ministry of the central government are not systematically linked together. In addition, horizontal relations between local governments and local educational authorities are not cooperatively operated. Recognizing these facts, this article analysed major problems and features of local education finance through the basic statistical analysis. Based on the results, this study proposed short-term and long-term policy proposals for the improvement of local education financing system. As the short and medium term policy proposal, this study proposed the following measures: (1) allowing partial taxing powers to local education authorities, (2) strengthening the linkage between the Local Shared Tax and the Local Shared Tax for Education, (3) strengthening the linkage between local government and local education authority in the areas of related policy and fund(budget) utilization.

IEC61850 기반 변전소자동화시스템의 신뢰도 향상기법

임성일(Seong-Il Lim) 한국조명·전기설비학회 2008 조명·전기설비학회논문지 Vol.22 No.9

최근 IEC61850이 변전소자동화시스템의 국제표준으로 자리 잡고 있는데, 그 중에서도 보호IED의 신뢰도는 매우 중요하다. 본 논문에서는 IEC61850의 특징을 이용하여 보호IED의 신뢰도를 향상시키는 새로운 방법을 제시한다. 본 논문에서 제시하는 신뢰도 향상기법은 적은 수의 백업 IED를 사용하므로 경제적이면서도, 기존의 2계열화나 백업보호방식에 비하여 오부동작을 현저히 감소시킬 수 있다. 6모선 예제계통에 대한 모의실험을 통하여 본 논문에서 제시하는 신뢰도 향상기법의 유용성을 검증하였다. The effect of the fault could be propagated to healthy region when the fault on the power system is not cleared in time. In order to enhance reliability of the protective relay, a dual and/or backup system is adopted for the power system protection. The IEC61850 has become an international standard of the substation automation system for interoperability between heterogeneous IEDs from different vendors. This paper introduces a reliability enhancement technique for the substation automation system using characteristics of the IEC61850.