꽃게(Portunus trituberculatus) 헤모시아닌의 CNBr-peptide에 관한 연구

김상복,유병선,이강석,Kim, Sang-Bog,Yoo, Byung-Sun,Lee, Kang-Suk 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.2

정제된 꽃게(Portunus trituberculatus) 헤모시아닌에 CNBr을 처리한 후 전기영동을 한 결과 분자량이 각각 31,000, 19,000, 13,000, 6,000 및 4,000인 다섯 개의 peptide로 절단되었다. 이들을 전기영동도에 따라서 peptide C1부터 C5로 각각 명명하였다. CNBr로 처리된 헤모시아닌을 Sephadex G-50에서 5% propionic acid로 용출할 경우 3개의 분획으로 분리되었고 이들 각 분획에 대한 전기영동 결과 첫번째 분획에는 peptide C1, C2, C3가 응집된 상태로 용출되었고, 두번째 분획에서는 peptide C4와 소량의 C2, C3가, 세번째 분획에서는 peptide C5가 각각 존재함을 확인하였다. Peptide C5는 $Cu^{++}$가 결합된채 분리되었다. Sephadex G-50 에서 용출된 각 분획을 DEAE-cellulose column chromatography를 이용하여 peptide C4와 peptide C5를 순수분리하였다. 순수분리된 peptide C4와 peptide C5 및 꽃게 헤모시아닌에 대한 아미노산 조성을 본 결과 꽃게 헤모시아닌은 Aspartic acid와 Glutamic acid 함량이 높은 것으로 밝혀졌다. Peptide C4의 경우는 몇 개 아미노산을 제외하고는 꽃게 헤모시아닌과 큰 차이가 없었으나 $Cu^{++}$를 함유하고 있는 peptide C5는 Lysine 함량이 특히 높은 강한 염기성으로 나타냈다. Purified hemocyanin of the crab, Portunus Trituberculatus was treated with CNBr. The SDS electrophoresis of CNBr-treated hemocyanin showed five distinct peptides fragment, referred to as peptide C1, C2, C3, C4, and C5, respectively, according to the mobility in electrophoresis gel. Molecular weights of each peptide estimated by SDS polyacrylamide gel electrophoresis were 31,000, 19,000, 13,000, 6,000, and 4,000, respectively. In order to separate peptides, CNBr-treated hemocyanin was eluted with 5% propionic acid on Sephadex G-50 column chromatography, in which three fractions were obtained. Fraction I contained peptide C1, peptide C2, peptide C3. Peptide C4 was eluted in fraction II with some impurity of peptide C2 and C3, and peptide C5 in fraction III, respectively. Peptide C5 was the smallest fragment and showed the copper-binding property. The amino acid compositions of the whole hemocyanin and CNBr fragments (peptide C4 and peptide C5) have been analyzed. The amino acid composition of whole hemocyanin showed that P. trituberculatus hemocyanin was typically acidic protein. While the amino acid composition of peptide C4 was similar to that of whole hemocyanin, peptide C5, containing copper, showed high content of lysine residue.

꽃게 체액에서 단백질 분해효소 억제물질의 정제

김상복,이강석,유병선,장정순,김정규,Kim, Sang-Bog,Lee, Kang-Suk,Yoo, Byung-Sun,Chang, Chung-Soon,Kim, Jung-Kyu 생화학분자생물학회 1993 한국생화학회지 Vol.26 No.6

꽃게(Portunus trituberculatus) 체액을 $Zn^{2+}$ -chelate affinity column을 이용하여 분리 정제하였다. 그 결과 척추동물의 경우와는 달리 affinity gel의 재생과정($Zn^{2+}$ 이온제거)인 50mM EDTA(ethylene diaminetetraacetic acid) 분획에서 ${\alpha}-macroglobulin({\alpha}-M)$이 분리되였다. 꽃게의 ${\alpha}-macroglobulin$은 trypsin, chymotrypsin, thermolysin의 활성을 억제하였으나, 사람의 ${\alpha}_{2}-macroglobulin$과는 달리 subtilisin과 cathepsin C 등의 활성은 억제하지 못하였다. 또한 고등동물의 경우와 달리 trypsin 보다는 thermolysin과의 친화력이 높았다. 꽃게 ${\alpha}-M$은 monomer의 분자량이 180 kDa인 dimer의 행태로 존재하며 상기 monomer는 열과 detergent(SDS)에 의해서 분자량 95 kDa 및 85 kDa의 polypeptide로 분리되었다. A high molecular weight proteinase inhibitor has been purified from the body fluid of sea crab (Portunus trituberculatus) using biospecific $Zn^{2+}$ chelate affinity chromatography. The proteinase inhibitor inhibits the proteolytic activity of a variety of endopeptidase such as trypsin, chymotrypsin and thermolysin. Like other vertebrate ${\alpha}-M$, the sea crab ${\alpha}-macroglobulin$ $({\alpha}-M)$ was composed of subunits of molecular weight 180 kDa as judged by polyacrylamide gel electrophoresis under reducing conditions. The apparent native molecular weight of the sea crab ${\alpha}-M$ determined by gel filtration was 360 kDa. Thermal fragmentation studies revealed that the monomer (180 kDa) was broken down into 95 kDa and 85 kDa fragments, respectively. On the basis of above results, it is concluded that the sea crab ${\alpha}-M$ is a dimer consisting of two weakly bound monomer,and the autolytic cleavage occurs easily by SDS and heat treatment.

Swamp eel ( Monopterus albus ) 혈장 Albumin 분획의 정제 및 물리화학적 성상에 관한 연구

김상복,장정순 ( Sang Bog Kim,Chung Soon Chang ) 생화학분자생물학회 1982 BMB Reports Vol.15 No.4

Albumin fraction was purified from the swamp eel by salting out with ammonium sulfate and following by DEAE-Sephadex A50 column chromatography. The purified albumin fraction appeared to be homogeneous by Davis and NaDodSO₄ polyacrylamide gel electrophoresis after albumin fraction was treated with organic cation, rivanol (2-ethoxy-6, 9, diaminoacridine lactate). The physico-chemical properties on purified albumin fraction were characterized by means of molecular weight determination, -SH content, extinction and sedimentation coefficient, etc.

PMO 수행 인력의 핵심 역량 중요도 비교 연구

김상복(Sang-Bog Kim),김진태(Jin-Tae Kim),장병만(Byung-Man Chang) 한국정보기술학회 2014 한국정보기술학회논문지 Vol.12 No.7

Recently as the importance of PMO increases, cases in which complaints about the result of the project and negative opinions were expressed have been reported due to lack of clear role establishment at the time of PMO introduction, inadequacy of functional definition and lack of competence on the part of performance staff. The (major) cause of these cases is lack of expertise of performing staffs. What is the core competence necessary to secure the expertise of PMO performance workforce? This study started from this question and, from existing cases, it drew out the competences that the performing staff should possess and then figured out core competences though fussy-based AHP analysis. After the drawing out of core competences, study was conducted on the difference of core competences by role of PMO staff. The essential significance of this study is that it provides important standards for the human resource development of PMO staff, required competence standards for mid-and-large-scale PMO projects and evaluation tool for the levels of PMO staff, and it developed education program to enhance the competence of PMO staff and has the potential to be applied to indexes when evaluations are conducted for commissioned PMO projects.

개인정보 영향평가 정보시스템 설계·구현 및 적용 사례

김상복(Sang-Bog Kim),장병만(Byung-Man Chang) 한국정보기술학회 2015 한국정보기술학회논문지 Vol.13 No.6

Privacy infringement accidents, which recently have occurred in various ways, show that they are man-made accidents and that they are confined to underdeveloped countries and that privacy is being very poorly managed. Privacy protection is not the exclusive duty of any one employee in an organization but an activity that requires all executives and employees to establish continuous improvement plans, implement them, and inspect related improvement tasks by raising awareness of privacy protection and diagnosing related activities accurately. Meanwhile, the reality is that small & medium-sized businesses are having trouble understanding privacy, recognizing risks associated with its management, and taking countermeasures. Therefore, this study is intended to suggest a management environment that ensures and promotes continuous privacy protection activities. The main focus of the study is to propose an environment that allows privacy operators to manage private information files with more ease and to have efficient means of communication that allows them to comply with the report form of PIA. In particular, Cloud services will be presented as a way to protect privacy effectively and policy directions for privacy protection will be suggested so that improvement plans can develop into common components. In addition, effective improvement tasks for privacy protection will be discussed with suggestions as to how they might be implemented.

정보시스템 총괄감리원과 수행감리원 간 핵심역량 중요도 비교 연구

김상복(Sang-Bog Kim),김승희(Seung-Hee Kim),김우제(Woo-Je Kim) 한국정보기술학회 2013 한국정보기술학회논문지 Vol.11 No.11

Even though the quality of information system auditor is recognized as an important factor for audit quality, there has been no study that addressed the issue about the auditors ability for each role. This study investigated the difference of abilities between general auditors and performance auditor by classifying auditors by performance role, producing the abilities that auditors should have based on existing cases and producing core abilities through AHP analysis, The analysis on the result showed that the essential abilities of general auditors are presentation ability, leadership, ability of strategic thinking and ability of communication with clients, while those of performance auditors are professional skill in auditing area and the ability of problem-solving. The result of this study is expected to provide a working-level implication to plan a training program for information system auditors such as training program of fostering auditors and maintenance training. It would also contribute to the improvement of auditing performance according to the auditing activities based on auditors ability.

꽃게 체액내 단백질 분해억제물질의 생화학적 특성

김상복,이강석,김인규,유병선,장정순,Kim, Sang-Bog,Lee, Kang-Suk,Kim, In-Kyu,Yoo, Byung-Sun,Chang, Chung-Soon 생화학분자생물학회 1993 한국생화학회지 Vol.26 No.8

꽃게(Portunus trituberculatus) 체액에서 정제된 ${\alpha}$-macroglobulin(${\alpha}$-M)은 단백질 분해억제물질로 고등동물의 ${\alpha}_2$-M과는 달리 trypsin과의 결합비는 casein 및 TAME를 기질로 사용하였을 때 1:0.3으로 닭, 개구리(Rana pipiens), 바다가재(Hamarus americanus) 등 보다 낮은 값을 나타냄으로써 하등으로 내려갈수록 trypsin과의 결합비가 점차 낮아진다는 사실을 알수 있었다. 한편 꽃게 ${\alpha}$-M을 methylamine으로 변성시킬 경우 trypsin의 활성은 억제되지 않았으며, 이러한 결과는 지금까지 밝혀진 척추 및 무척추동물의 ${\alpha}_2$-M 의 생화학적 특성과 일치하였다. 꽃게 ${\alpha}$-M의 아미노산 조성을 지금까지 밝혀진 다른 동물의 ${\alpha}_2$-M과 비교한 결과 aspartic acid의 함량이 다소 높은 것을 제외하고는 대부분의 아미노산 조성이 서로 비슷한 경향을 보였다. 이러한 꽃게 ${\alpha}$-M의 아미노산 조성을 근거로 $S{\Delta}Q$값을 사용하여 단백질의 근연관계를 조사한 결과 꽃게 ${\alpha}$-M은 개구리, 투구게(Limulus polyphemus)의 ${\alpha}$-M과 매우 가까운 단백질로 밝혀졌다. Trypsin binding studies showed that sea crab ${\alpha}$-macroglobulin (${\alpha}$-M) could bind 0.3 mol of trypsin per molecule. It can, therefore, be concluded that the molecular ratio of proteinase/inhibitor was different from the stoichiometry for human ${\alpha}_2$-M (half molecule) and also different from other data for the southern grass frog and American lobster where the values were in the range of 0.45~0.79 mol of trypsin/mol of macroglobulin, indicating the lower animals showed low trypsin binding capacity compared with that of higher animals. The ${\alpha}$-M of the sea crab completely lost its ability for inhibition of trypsin activity by methylamine. The analysis of $S{\Delta}Q$ parameter based on the amino acid compositions indicated that the sea crab ${\alpha}$-M is closely related with that of the frog (Rana pipiens) and the horseshoe crab, respectively.

꽃게 체액에서 단백질 분해효소 억제물질의 정제

김상복,이강석,유병선,장정순,김정규 ( Sang Bog Kim,Kang Suk Lee,Byung Sun Yoo,Chung Soon Chang,Jung Kyu Kim ) 생화학분자생물학회 1993 BMB Reports Vol.26 No.6

A high molecular weight proteinase inhibitor has been purified from the body fluid of sea crab (Portunus trituberculatus) using biospecific Zn^(2+) chelate affinity chromatography. The proteinase inhibitor inhibits the proteolytic activity of a variety of endopeptidase such as trypsin, chymotrypsin and thermolysin. Like other vertebrate α-M, the sea crab α-macroglobulin (α-M) was composed of subunits of molecular weight 180 kDa as judged by polyacrylamide gel electrophoresis under reducing conditions. The apparent native molecular weight of the sea crab α-M determined by gel filtration was 360 kDa. Thermal fragmentation studies revealed that the monomer (180 kDa) was broken down into 95 kDa and 85 kDa fragments, respectively. On the basis of above results, it is concluded that the sea crab α-M is a dimer consisting of two weakly bound monomer,and the autolytic cleavage occurs easily by SDS and heat treatment.

꽃게 체액내 단백질 분해억제물질의 생화학적 특성

김상복,이강석,김인규,유병선,장정순 ( Sang Bog Kim,Kang Suk Lee,In Kyu Kim,Byung Sun Yoo,Chung Soon Chang ) 생화학분자생물학회 1993 BMB Reports Vol.26 No.8

Trypsin binding studies showed that sea crab α-macroglobulin (α-M) could bind 0.3 ㏖ of trypsin per molecule. It can, therefore, be concluded that the molecular ratio of proteinase/inhibitor was different from the stoichiometry for human α₂-M (half ㏖ecule) and also different from other data for the southern grass frog and American lobster where the values were in the range of 0.45∼0.79 ㏖ of trypsin/㏖ of macroglobulin, indicating the lower animals showed low trypsin binding capacity compared with that of higher animals. The a-M of the sea crab completely lost its ability for inhibition of trypsin activity by methylamine. The analysis of SΔQ parameter based on the amino acid compositions indicated that the sea crab α-M is closely related with that of the frog (Rana pipiens) and the horseshoe crab, respectively.

꽃게 ( Portunus trituberculatus ) 헤모시아닌의 CNBr - peptide에 관한 연구

김상복,유병선,이강석 ( Sang Bog Kim,Byung Sun Yoo,Kang Suk Lee ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.2

Purified hemocyanin of the crab, Portunus Trituberculatus was treated with CNBr. The SDS electrophoresis of CNBr-treated hemocyanin showed five distinct peptides fragment, referred to as peptide C1, C2, C3, C4, and C5, respectively, according to the mobility in electrophoresis gel. Molecular weights of each peptide estimated by SDS polyacrylamide gel electrophoresis were 31,000, 19,000, 13,000, 6,000, and 4,000, respectively. In order to separate peptides, CNBr-treated hemocyanin was eluted with 5% propionic acid on Sephadex G-50 column chromatography, in which three fractions were obtained. Fraction I contained peptide Cl, peptide C2, peptide C3. Peptide C4 was eluted in fraction II with some impurity of peptide C2 and C3, and peptide C5 in fraction III, respectively. Peptide C5 was the smallest fragment and showed the copper-binding property. The amino acid compositions of the whole hemocyanin and CNBr fragments (peptide C4 and peptide C5) have been analyzed. The amino acid composition of whole hemocyanin showed that P. trituberculatus hemocyanin was typically acidic protein. While the amino acid composition of peptide C4 was similar to that of whole hemocyanin, peptide C5, containing copper, showed high content of lysine residue.