• BMB Reports
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• 제28권2호
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• pp.162-169
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• 1995

Rat brain succinic semialdehyde dehydrogenase (EC 1.2.1.24 SSADH) activity was detected in mitochondrial, cytosolic and microsomal fractions. Brain mitochondrial soluble SSADH was purified by ammonium sulfate precipitation, DEAE Sephacel, and 5'-AMP Sepharose 4B affinity chromatography. The purified enzyme was shown to consist of four identical subunits, and the molecular weight of a subunit was 55 kD. The $K_m$ for short chain aliphatic aldehydes and aromatic aldehydes were at the $10^{-3}M$ level but that for succinic semialdehyde was 2.2 ${\mu}M$. Either $NAD^+$ or $NADP^+$ can be used as a cofactor but the affinity for $NAD^+$ was 10 times higher than that for $NADP^+$. The brain cytosolic SSADH was also purified by ammonium sulfate precipitation, DEAE Sephacel, Blue Sepharose CL-6B and 5'-AMP Sepharose 4B affinity chromatography and its Km for short chain aliphatic aldehydes was at the $10^{-3}$ level but that for succinic semialdehyde was 3.3 ${\mu}M$. $NAD^+$ can be used as a cofactor for this enzyme. We suppose that both enzyme might participate in the oxidation of succinic semialdehyde, which is produced during GABA metabolism. The activity of both cytosolic and mitochondrial SSADH was markedly inhibited when the concentration of succinic semialdehyde was high. The reciprocal plot pattern of product inhibition and initial velocity indicated a sequential ordered mechanism for mitochondrial matrix SSADH. Chemical modification data suggested that amino acid residues such as cysteine, serine and lysine might participate in the SSADH reaction.

• Bulletin of the Korean Chemical Society
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• 제15권5호
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• pp.394-399
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• 1994

The catalytic mechanism of malonyl-CoA synthetase from Rhizobium trifolii was investigated by the steady state kinetics and intermediate identification. Initial velocity studies and the product inhibition studies with AMP and PPi strongly suggested ordered Bi Uni Uni Bi Ping-Pong Ter Ter system as the most probable steady state kinetic mechanism of malonyl-CoA synthetase. Michaelis constants were $0.17{\pm}0.04 {\mu}M,\;0.24{\pm}0.18 {\mu}M\;and\;0.045{\pm}0.26 {\mu}$M for ATP, malonate and CoA, respectively. The TLC analysis of the $^{32}P-labelled$ products in reaction mixture containing $[{\gamma}-^{32}P]$ ATP in the absence of CoA showed that PPi was produced after the sequential addition of ATP and malonate. Formation of malonyl-AMP, suggested as an intermediate in the kinetically deduced mechanism, was confirmed by the analysis of $^{31}P-NMR$ spectra of AMP product isolated from the $^{18}O$ transfer experiment using $[^{18}O]$malonate. Two resonances were observed, corresponding to AMP labelled with zero and one atom of $^{18}O$, indicating that one atom of $^{18}O$ transferred from $[^{18}O]$malonate to AMP through the formation of malonyl-AMP. Formation of malonyl-AMP was also confirmed through the TLC analysis of reaction mixture containing $[{\alpha}-^{32}P]$ATP. These results strongly support the ordered Bi Uni Uni Bi Ping-Pong Ter Ter mechanism deduced from the initial velocity and product inhibition studies.

• 인터넷정보학회논문지
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• 제12권4호
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• pp.35-43
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• 2011

최근 들어 무선인지 애드 혹 네트워크(CRAHN)는 임시적으로 활용되지 않은 라이선스 스펙트럼을 기회주의적 방법으로 사용하여 통신하는 차세대 애드 혹 네트워크로 각광받고 있다. 여타 무선인지 시스템에서와 같이, 정확한 스펙트럼 센싱은 무선인지 애드 혹 네트워크 구현을 위한 필수 요소 기술이다. 중앙 제어국 중심의 협력 스펙트럼 센싱 기법은 센싱 성능을 향상 시킬 수 있으나, 무선인지 애드 혹 네트워크와 같은 비인프라망에서는 중앙 제어국이 없어 협력 센싱 구현이 어렵다. 본 논문에서는 무선인지 애드 혹 네트워크를 위한 새로운 순차적 협력 스펙트럼 센싱 기법을 제안한다. 제안된 기법에서는 Dempster Shafer 이론 기반으로 국부 스펙트럼 센싱 데이터의 신뢰도가 가장 높은 인지 노드가 임시 중앙국이 되어 주변 센서 노드로부터 스펙트럼 센싱값을 수집하여 스펙트럼 사용 유무에 대한 최종 결정을 만들고 그 최종 결정을 이웃 노드들에게 전송한다. 또한 본 논문에서는 분산 방법으로 개별 인지 노드들이 자신의 센싱 데이터를 효율적으로 교환하기 위한, 센싱데이터 신뢰도 순서 기반의 전송 메커니즘을 제안하였다. 시뮬레이션을 통해 제안된 방식은 기존 중앙 제어국 중심의 협력 스펙트럼 센싱 기법과 유사한 스펙트럼 센싱 성능을 제공하는 동시에, 최종 스펙트럼 결정까지 소요되는 센싱 데이터 수집시간 및 교환되는 센싱 데이터 량을 현저히 줄일 수 있음을 보였다.

• Bulletin of the Korean Chemical Society
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• 제23권8호
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• pp.1057-1061
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• 2002

The Hafnia alvei aspartase activity with acetic anhydride treatment gradually increased and reached 7.5-fold that of the native one. The activity of the acetylated aspartase was a little higher than that of the native enzyme, indicating that the cooperativity between a substrate and enzyme is increased. The optimum temperature of the native asparatse was $45^{\circ}C$, and that of the acetylated enzyme shifted to $40^{\circ}C.$ The pH vs. the activity profile of the acetylated asparatse was also different from that of the native enzyme. The initial velocity pattern of the acetylated aspartase intersects to the left of the ordinate, indicating the sequential kinetic mechanism other than a rapid equilibrium ordered one. The reciprocal plots for aspartate of the native aspartase were curved, but those of the acetylated aspartase were linear, indicating the Michaelis-Menten kinetics. The helical content of the acetylated aspartase was rather decreased to $9{\textperthousand}$ than that $(63{\textperthousand})$ of the native one.