Essential Arginyl Residue(s) of Glycolate Oxidase from Rat Liver

최정도,Choi, Jung-Do,Horiike, Kihachiro,Nozaki, Mitsuhiro 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.3

쥐 간으로부터 분리한 glycolate oxidase는 arginine과 선택적으로 반응하는 시약, phenylglyoxal이나 2, 3-butadione과의 반응에 의하여 효소 활성을 상실하였다. Phenylglyoxal에 의한 효소의 비활성화 반응은 유사 일차반응, phenylglyoxal에 대해서는 일차반응 이였으며, 장시간 반응에 의해서 효소는 완전히 비활성화 되었다. 기질 glycolate나 경쟁적 저해제 phenyllactic acid와 oxalate는 효소의 비활성화에 대해 방어효과를 보여주었다. 다른 종류의 dicarbonyl 화합물 butanedione도 효소와 반응하여 효소를 비활성화 시키나, 반응속도는 phenylglyoxal 반용에 비해 매우 느린 편이다. Butanedione에 의한 효소의 비활성화 반응의 속도도 반응시약의 농도에 비례하였으며, borate 용액은 반응속도를 증가시키는 효과를 나타내었다. Phenylglyoxal이나 butanedione과의 반응에 의해 활성이 감소된 효소용액으로부터 구한 glycolate의 Km 값은 대조 실험으로부터 얻은 Km값과 유사하였다. 아미노산 분석에 의하여 변형된 arginine의 정확한 숫자는 알 수 없었으나, phenylglyoxal에 의하여 80% 정도 활성을 상실한 효소는 소단위 내의 15개의 arginine중에서 1-3개를 상설했으며, 경쟁적 저해제는 arginine 변형에 대해서도 방어효과를 나타내었다. 이러한 실험결과로부터, 효소의 활성부위의 arginine이 기질의 결합에 중요한 역할을 하는 것으로 추측된다. Glycolate oxidase from rat liver is inactivated by arginine-specific reagents, phenylglyoxal and 2,3-butanedione. Inactivation of the oxidase by phenylglyoxal follows pseudo-first order kinetics, and is first order with respect to the reagent concentration. The oxidase is completely inactivated by prolonged incubation with phenylglyoxal. The substrate glycolate and substrate-competitive inhibitors, phenyllactic acid and oxalate protect the oxidase against inactivation. Other dicarbonyl compound butanedione also reacts with the enzyme to cause loss of activity but at much slower rate. Inactivation by butanedione is also dependent on the reagent concentration, and enhanced by the presence of borate buffer. Kinetic studies with the enzyme partially inactivated by phenylglyoxal or butanedione demonstrate that Km value of glycolate is similar to the constant for the control enzyme. Although amino acid analysis data do not provide the precise number of arginine modified, a few arginyl residues out of the 15 total residues in the subunit seem to be modified upon 80% inactivation by phenylglyoxal, and competitive inhibitor provides some protection against modification of arginyl residues. The present data suggest that arginyl residue(s) is probably involved in the binding of substrate to the enzyme.

Absorption and Fluorescence spectroscopic Studies of Glycolate Oxidase

최정도,박금수,양덕조,Choi, Jung-Do,Park, Kum-Soo,Yang, Duk-Jo Korean Society for Biochemistry and Molecular Biol 1990 한국생화학회지 Vol.23 No.3

Glycolate oxidase의 활성부위의 성질을 규명하기 위하여 흡광 및 형광의 분광학적 연구를 수행하였다. 효소에 결합한 FMN의 전자전이 스펙트럼은 300-500nm 영역에서 450nm과 340nm에 정점을 가진 두 개의 띠를 갖고 있었다. 모델계의 흡수 스펙트라와 비교한 결과, 효소에 결합한 FMN은 거의 수소결합을 하지 않은 상태로 존재하며, 주위 환경은 물보다 훨씬 극성이 낮은 것으로 추측된다. 효소에 결합한 FMN의 형광은 비결합 FMN 형광의 5% 정도이며, 낮은 형광 수율은 FMN과 단백질내의 방향성 아미노산과 강력한 작용이 있음을 시사한다. 비결합 FMN과 결합 FMN의 형광의 ?봬た? 관한 연구결과로 부터 조효소 FMN은 표면에 노출되어 있지 않고 효소내의 좁은 틈 사이에 존재하며 수화상태의 큰 이온이 접근하기 어려운 것으로 추측된다. 효소 분자를 구성하는 각 단위내에 한 개의 cysteine이 존재하며, 이 cysteine에 은 이온의 접근이 가능하며, 이 아미노산은 효소의 활성부위에 위치하지 않고 그 근처에 있을 것으로 생각된다. Absorption and fluorescence spectroscopic studies of glycolate oxidase were carried out to elucidate the properties of the active site of the enzyme. The electronic spectrum of FMN bound to the enzyme consists of two bands centered around 450 nm and 340 nm in the region of 300-500 nm. Comparing with the spectra of model systems, it is suggested that the bound FMN existes almost as non-hydrogen-bounded species, and the environment of the FMN is much less polar than water medium. The fluorescence of the bound FMN is only about 5% as great as that of free FMN indicating strong interactions between flavin moiety and aromatic amino acid residues of the protein. The fluorescence quenching data of free FMN and bound FMN suggest that the coenzyme FMN is not exposed on the surface and most likely located in a crevice whose dimensions are not large enough for large hydrated ions to enter it. The cysteine residue, a subunit of the enzyme is known to have one cysteine, is accesible to silver ion and probably located not at active site but near active site.

Glycolate Oxidase 에 관한 흡광 및 형광의 분광학적 연구

최정도,박금수,양덕조 ( Jung Do Choi,Kum Soo Park,Duk Jo Yang ) 생화학분자생물학회 1990 BMB Reports Vol.23 No.3

Absorption and fluorescence spectroscopic studies of glycolate oxidase were carried out to elucidate the properties of the active site of the enzyme. The electronic spectrum of FMN bound to the enzyme consists of two bands centered around 450 nm and 340 nm in the region of 300-500 nm. Comparing with the spectra of model systems, it is suggested that the bound FMN existes almost as non-hydrogen-bounded species, and the environment of the FMN is much less polar than water medium. The fluorescence of the bound FMN is only about 5% as great as that of free FMN indicating strong interactions between flavin moiety and aromatic amino acid residues of the protein. The fluorescence quenching data of free FMN and bound FMN suggest that the coenzyme FMN is not exposed on the surface and most likely located in a crevice whose dimensions are not large enough for large hydrated ions to enter it. The cysteine residue, a subunit of the enzyme is known to have one cysteine, is accesible to silver ion and probably located not at active site but near active site.

The Interaction of D-Amino Acid Oxidase with Carboxylic Acid Inhibitors

최정도,Choi, Jung-Do 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.2

많은 종류의 카르복실산은 D-amino acid oxidase와 복합체 (complex)를 형성함으로 효소의 흡광과 CD 스펙트럼 그리고 형광에 영향을 미친다. 복합체 형성에 의해 350-500 nm 범위에서 흡광 스펙트럼의 변화가 일어나며, 이 변화는 조효소 FAD의 isoalloxazine 환에 있는 질소나 산소 원자에 형성되는 수소결합에 의한 영향으로 어느 정도 설명된다. 카르복실산과의 복합체 형성으로 효소의 370 nm와 450 nm의 CD band의 강도가 증가하는데, 이 변화는 FAD의 isoalloxazine 환과 ribityl 고리와의 상호작용의 변화에 기인된 것으로 생각된다. 효소와 저해제 사이에 복합체를 형성할 경우 조효소 FAD의 형광이 현저히 감소되는데 이는 flavin 핵과 저해제와의 직접적인 상호작용에 의한 결과로 추측된다. 완전효소와 효소-저해제 복합체내에 있는 FAD의 형광의 quenching에 관한 연구 결과는 조효소 FAD가 효소의 표면에 노출되어 있지 않으며, 효소 단백질과 FAD 결합에 소수성 상호작용과 정전기적 인력이 기여하고 있을 것임을 시사했다. A number of carboxylic acids affect the absorption and circular dichroism spectra and fluorescence of D-amino acid oxidase by formation of complexes with the enzyme. The absorption spectral change in the 350 to 500 nm region upon complex formation could be interpreted at least in part as the effects of hydrogen bonding at nitrogen and oxygen atoms of isoalloxazine moiety of coenzyme flavin adenine dinucleotide(FAD). The effect of complex formation in intensifying both CD bands of 370 and 450 nm regions is discussed in terms of the interaction of ribityl chain with isoalloxazine ring of FAD. A marked fluorescence quenching of FAD chromophore in the enzyme-inhibitor complexes is probably due to direct interaction between the flavin nucleus and inhibitor. The fluorescence quenching studies of FAD in holoenzyme and its complexes indicate that the coenzyme FAD is not exposed on the enzyme surface, and both hydrophobic and electrostatic interactions contribute to the binding of FAD to the apoenzyme.

쥐 간의 Glycolate Oxidase 의 필수 Arginine

최정도,Kihachiro Horiike,Mitsuhiro Nozaki ( Jung Do Choi,Kihachiro horiike . Mitsuhiro Nozaki ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.3

Glycolate oxidase from rat liver is inactivated by arginine-specific reagents, phenylglyoxal and 2,3-butanedione. Inactivation of the oxidase by phenylglyoxal follows pseudo-first order kinetics, and is first order with respect to the reagent concentration. The oxidase is completely inactivated by prolonged incubation with phenylglyoxal. The substrate glycolate and substrate-competitive inhibitors, phenyllactic acid and oxalate protect the oxidase against inactivation. Other dicarbonyl compound butanedione also reacts with the enzyme to cause loss of activity but at much slower rate. Inactivation by butanedione is also dependent on the reagent concentration, and enhanced by the presence of borate buffer. Kinetic studies with the enzyme partially inactivated by phenylglyoxal or butanedione demonstrate that Km value of glycolate is similar to the constant for the control enzyme. Although amino acid analysis data do not provide the precise number of arginine modified, a few arginyl residues out of the 15 total residues in the subunit seem to be modified upon 80% inactivation by phenylglyoxal, and competitive inhibitor provides some protection against modification of arginyl residues. The present data suggest that arginyl residue(s) is probably involved in the binding of substrate to the enzyme.

추구편익에 따른 대학생들의 응급처치 및 심폐소생술 교육프로그램 만족도, 재방문의도 및 추천의도 간의 관계

최정도(Jung Do Choi),문태영(Tae Young Moon) 한국사회체육학회 2009 한국사회체육학회지 Vol.0 No.38

The purpose of this study was to examine the satisfaction level of participants in educational programs of first-aid treatment and cardiopulmonary resuscitation with the programs, their behavioral intention and the relationship of variables related to the satisfaction level and revisit intention and recommendation in an effort to provide information to first-aid treatment and CPR educators and people concerned. The subjects in this study were 300 students who participated in first-aid and CPR education programs. After a survey was conducted, the collected data were analyzed with SPSS 14.0 and AMOS 16.0 programs. Descriptive statistics, factor analysis, reliability analysis and structural equation model were utilized. The findings of the study were as follows: The convenience pursuit of the participators had a significant impact on their satisfaction level with the educational programs and behavioral intention, and all the hypotheses formulated in the study were accepted. First-aid and CPR educators and people concerned should provide more systematic first-aid and CPR education, and prepare a variety of programs in order for many students to receive the education.

스쿠버다이빙 소비자의 참여정도, 라이프스타일 및 만족도의 관계

최정도 ( Jung Do Choi ) 한국스포츠산업경영학회 2007 한국스포츠산업경영학회지 Vol.12 No.4

조건기반의 지능형 홈 오토메이션 시스템

최정도(Jung-Do Choi),이주엽(Ju-Youb Lee),강수훈(Jung-Do Choi),박진희(Ju-Youb Lee),신동석(Su-Hoon Kang) 한국컴퓨터정보학회 2009 한국컴퓨터정보학회 학술발표논문집 Vol.16 No.2

현재 홈 네트워크 환경은 각 디바이스들을 위한 독립적인 서비스 보다 모든 디바이스를 통합관리 지원하는 지능적인 홈 오토메이션 서비스로 발전하고 있다. 이에 본 논문에서는 사용자의 신원확인만으로 댁내의 모든 디바이스들을 사용자 개인의 특성에 맞게 자동 제어할 수 있는 지능형 홈 오토메이션 시스템을 제안한다. 등록된 사용자의 정의된 규칙을 이용하여 가정내의 모든 디바이스들을 자동 제어한 수 있는 조건기반엔진을 설계하여 모든 디바이스를 독립적으로 제어하는 방식을 보완함으로써 지능적인 서비스를 사용자에게 제공할 수 있다.

D - Amino acid oxidase 와 카르복실산 저해제와의 상호작용

최정도 ( Jung Do Choi ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.2

A number of carboxylic acids affect the absorption and circular dichroism spectra and fluorescence of D-amino acid oxidase by formation of complexes with the enzyme. The absorption spectral change in the 350 to 500 nm region upon complex formation could be interpreted at least in part as the effects of hydrogen bonding at nitrogen and oxygen atoms of isoalloxazine moiety of coenzyme flavin adenine dinucleotide(FAD). The effect of complex formation in intensifying both CD bands of 370 and 450 nm regions is discussed in terms of the interaction of ribityl chain with isoalloxazine ring of FAD. A marked fluorescence quenching of FAD chromophore in the enzyme-inhibitor complexes is probably due to direct interaction between the flavin nucleus and inhibitor. The fluorescence quenching studies of FAD in holoenzyme and its complexes indicate that the coenzyme FAD is not exposed on the enzyme surface, and both hydrophobic and electrostatic interactions contribute to the binding of FAD to the apoenzyme.

공공스포츠센터의 서비스품질과 고객만족 및 선택행동의 관계

최정도(Jung Do Choi),문태영(Tae Young Moon) 한국사회체육학회 2008 한국사회체육학회지 Vol.0 No.34

This study closely examines relationship among the service quality, customer satisfaction and choosing behavior in public sports center, thereby aiming to solve problems in light of operation that the public sports center has now, and to offer basic data for activation and efficient operation in the public sports center down the road. Regarding the subjects in this study, 271 materials were used for the final analysis. The collected data was analyzed by using SPSS 14.0 for Windows. Reliability analysis and factor analysis were carried out. T-test, ANOVA, Duncan`s post-test, Multiple Regression Analysis, and hierarchical regression analysis were performed. The following are the conclusions that were obtained through the above research method and procedure. First, the service quality in public sports center, re-visit intention, and recommendation intention were indicated to have difference depending on demographic characteristic variables. Second, there is relationship between the service quality of public sports center and revisit intention & recommendation intention. Third, the service quality of public sports center was indicated to have effect on customer satisfaction, revisit intention, and recommendation intention.