• Title/Summary/Keyword: Binding time

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Time-resolved Fluoroimmunoassay for the Measurement of 17$\beta$-Estradiol using Anti-idiotypic Antibody (Anti-idiotype 항체를 이용한 17$\beta$-Estradiol 측정을 위한 Time-resolved Fluoroimmunoassay)

  • 김윤규;김창규;박성민;이치호;이원창;최영숙;김종배
    • Korean Journal of Animal Reproduction
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    • v.16 no.4
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    • pp.325-333
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    • 1993
  • A competitive type immunoassay method for 17$\beta$-estradiol(E2) based on the idiotypic anti-idiotypic antibody and time-resolved fluorescence is described. The anti-idiotypic antibody(Ab2) produced to E2 binding site of the primary idiotype antibody (Ab1) was labelled with europium and was allowed to compete with E2 standards or serum sample for the binding sites of Ab1 which was bound to 2nd antibody captured ontothe surface of microtitre plates. Fluorescence measured by time-resolved fluorometer was inversely proportional to the concentration of E2 over the range 5~500pg/well. The sensitivity of the assay was 5pg per well which was compatible with that ofradioimmunoassay using the same Ab1 and 3H-E2 as a tracer. One great advantage of this method described here was to enable antibodies to be labelled instead of haptens, and thus makes it easier to develop sensitive and robust immunoassay systems specially for haptens.

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Conformational Dynamics of Heme Pocket in Myoglobin and Hemoglobin

  • Kim, Seong-Heun;Heo, Jeong-Hee;Lim, Man-Ho
    • Bulletin of the Korean Chemical Society
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    • v.26 no.1
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    • pp.151-156
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    • 2005
  • The conformational dynamics of heme pocket, a small vacant site near the binding site of heme proteins -myoglobin (Mb) and hemoglobin (Hb), was investigated after photolysis of carbon monoxide from MbCO and HbCO in D$_2$O solution at 283 K by probing time-resolved vibrational spectra of photolyzed CO. Two absorption bands, arising from CO in the heme pocket, evolve nonexponentially in time. The band at higher energy side blue shifts and broadens with time and the one at lower energy side narrows significantly with a negligible shift. These spectral evolutions are induced by protein conformational changes following photolysis that modify structure and electric field of heme pocket, and ligand dynamics in it. The conformational changes affecting the spectrum of photolyzed CO in heme pocket likely modulates ligand-binding activity.

The Slow and Tight Binding of MR-387A to Aminopeptidase N

  • CHUNG, MYUNG-CHUL;HYO-KON CHUN;HO-JAE LEE;CHOONG-HWAN LEE;SU-IL KIM;YUNG-HEE KHO
    • Journal of Microbiology and Biotechnology
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    • v.6 no.4
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    • pp.250-254
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    • 1996
  • MR-387A [(2S, 3R)-2-hydroxy-3-amino-4-phenylbutanoyl-L-valyl-L-prolyl-(2, 4-trans)- L-4-hydroxy-proline] reversibly inhibits aminopeptidase N (BC 3.4.11.2) in a process that is remarkable for its unusual degree of time dependence. The time required to inactivate the enzyme by 50$%$ ($t_{1/2}$) for establishing steady-state levels of $EI^*$complex was approximately 5 minutes. This indicates that the inhibition is a slow-binding process. In dissociation experiments of $EI^*$ complex, enzymic activity was regained slowly in a quadratic equation, indicating that the inhibition of aminopeptidase N by MR-387A is tight-binding and reversible. Thus, the binding of MR-387A by aminopeptidase N is slow and tight, with $K_{i}$ (for initial collision complex, EI) and $K_i{^*}$ (for final tightened complex, $EI^*$) of $2.2\times10^{-8}$ M (from Lineweaver-Burk plot) and $4.4\times10^{-10}$ M (from rate constants), respectively. These data indicate that MR-387A and aminopeptidase N are bound approximately 200-fold more tightly in the final $EI^*$complex than in the initial collision EI complex.

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A Construction of TMO Object Group Model for Distributed Real-Time Services (분산 실시간 서비스를 위한 TMO 객체그룹 모델의 구축)

  • 신창선;김명희;주수종
    • Journal of KIISE:Computer Systems and Theory
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    • v.30 no.5_6
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    • pp.307-318
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    • 2003
  • In this paper, we design and construct a TMO object group that provides the guaranteed real-time services in the distributed object computing environments, and verify execution power of its model for the correct distributed real-time services. The TMO object group we suggested is based on TINA's object group concept. This model consists of TMO objects having real-time properties and some components that support the object management service and the real-time scheduling service in the TMO object group. Also TMO objects can be duplicated or non-duplicated on distributed systems. Our model can execute the guaranteed distributed real-time service on COTS middlewares without restricting the specially ORB or the of operating system. For achieving goals of our model. we defined the concepts of the TMO object and the structure of the TMO object group. Also we designed and implemented the functions and interactions of components in the object group. The TMO object group includes the Dynamic Binder object and the Scheduler object for supporting the object management service and the real-time scheduling service, respectively The Dynamic Binder object supports the dynamic binding service that selects the appropriate one out of the duplicated TMO objects for the clients'request. And the Scheduler object supports the real-time scheduling service that determines the priority of tasks executed by an arbitrary TMO object for the clients'service requests. And then, in order to verify the executions of our model, we implemented the Dynamic Binder object and the Scheduler object adopting the binding priority algorithm for the dynamic binding service and the EDF algorithm for the real-time scheduling service from extending the existing known algorithms. Finally, from the numerical analyzed results we are shown, we verified whether our TMO object group model could support dynamic binding service for duplicated or non-duplicated TMO objects, also real-time scheduling service for an arbitrary TMO object requested from clients.

Effects of Thyroid Hormone on Pteroylpolyglutamate Chain Length and the Binding Activity of Folate Binding Protein in Rat Liver (갑상선 호르몬이 흰쥐 간세포내 엽산의 Polyglutamate 직쇄분포와 세포질 엽산 결합단백질의 결합성에 미치는 영향)

  • 민혜선
    • Journal of Nutrition and Health
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    • v.32 no.4
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    • pp.369-375
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    • 1999
  • Hyperthroidism in known to alter the activity of a number of enzymes involved in the catabolism of histidine to CO2. 10-Formyltetrahydrofolate dehydrogenase(EC 1.5, 1.6, 10-formyl-THE dehydrogenase) catalyzes the NADP-dependent conversion of 10-formyltetrahydrofolate to tetrahydrofolate and CO2. In previous studies, 10-formyl-THF dehydrogenase purified from rat and pig liver was coidentified with the cytosolic folate-binding protein. In this study, we investigated the effects of feeding thyroid powder (TP) and thiouracil(TU) on the folate-binding properties of 10-formyl-THE dehydrogenase, the uptake of an injected dose of [3H] folate, and the metabolism of labeled folate to pteroylopoly-${\gamma}$-glutamate in rat liver. The initial hepatic uptake(24hr) of the labeled folate dose was higher in TU-rats and slightly higher in TP-rats in controls. With longer time periods, decreased hepatic uptake of labeled folate was observed in TP-animals compared to euthroid animals, and high levels of hepatic uptake of labeled folate were maintained in TU-animals. This data shows that high levels of thyroid hormone decreased the retention of folate in rat liver. Folate polygutamate chain length was shorter in TU-rats than controls, which suggests that thyroid states do not affect the ability to synthesize pteroylpolyglutamates and that folate polyglutamate might be modulated by altered folate pool size. The ability of 10-formyl-THE dehydrogenase to bind folate in rat liver was similar in both TP-and TU-rats although dehydrogenase activity was changed by thyroid sates.

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An Immobilization of Extracellular Laccase to Humus-Iron Complex

  • Ginalska, Grazyna;Cho, Nam-Seok;Lobarzewski, Jerzy;Piccolo, Alessandro;Leonowicz, Andrzej
    • Journal of the Korean Wood Science and Technology
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    • v.29 no.3
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    • pp.104-111
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    • 2001
  • There are some evidence that active enzymatic proteins, e.g. fungal laccase, exist in the naturally occured soil humus. This study was performed to investigate the covalent binding of fungal laccase to the humic acid-iron complex, and to measure laccase activity of immobilized ones. Seven methods were adopted to form the covalent binding of fungal laccase with soil humic acids complexed with iron. Using these seven methods it was possible to change the dimension of spacer arm between laccase and support, and also to regulate the mode of covalent binding of this enzyme. The spacer arm was regulated from 2C to 11C. There was not observed any straight relationship between the spacer arm longitude and the laccase activity after immobilization, but the binding mode more effective than the former. Three out of the seven methods gave the high activity of immobilized laccase, and which active products of laccase immobilization was stable up to 10 days after the process. It is indicated that natural soil condition might be prevented the laccase activation by the toxic influence of some phenolic humic compounds. It was shown, for the first time, the possibilities to obtain the high activity of fungal laccase by binding to humic acids, and especially in complex with iron.

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A Remote SNMP Connection Request Mechanism for NATed Devices using UDP Hole Punching and Heuristic Hole Binding Time Search (UDP 홀 펀칭과 경험적 홀 유지시간 탐색을 이용한 NAT 환경단말의 SNMP 원격 접속요청 메커니즘)

  • Park, Choon-Gul;Kim, Seong-Il;Jeong, Ki-Tae;Lee, Young-Seok
    • Journal of KIISE:Information Networking
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    • v.35 no.5
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    • pp.367-373
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    • 2008
  • Recently, the NAT middlebox widely deployed in the home network environment prohibits DM operations from reaching user devices behind NAT. In this article, we focus on NAT issues to manage home network devices. Particularly, we discuss standardization efforts, and present our proposal to deploy DM services for VoIP and IPTV devices under NAT. By slightly changing behaviors of Simple Network Management Protocol (SNMP) Manager and Agent, and defining additional Management Objects (MOs) to gather NAT binding information, we could solve the NAT traversal problem under symmetric NAT. Moreover, we propose an enhanced method to search the UDP hole binding time of the NAT box. We applied our method to randomly selected 22 VoIP devices out of 194 NATed hosts in the real broadband network and have achieved 99% of the success ratio for exchanging SNMP request messages and 26% of enhancement for searching the UDP hole binding time.

Sleep Promoting Effect of Luteolin in Mice via Adenosine A1 and A2A Receptors

  • Kim, Tae-Ho;Custodio, Raly James;Cheong, Jae Hoon;Kim, Hee Jin;Jung, Yi-Sook
    • Biomolecules & Therapeutics
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    • v.27 no.6
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    • pp.584-590
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    • 2019
  • Luteolin, a widespread flavonoid, has been known to have neuroprotective activity against various neurologic diseases such as epilepsy, and Alzheimer's disease. However, little information is available regarding the hypnotic effect of luteolin. In this study, we evaluated the hypnotic effect of luteolin and its underlying mechanism. In pentobarbital-induced sleeping mice model, luteolin (1, and 3 mg/kg, p.o.) decreased sleep latency and increased the total sleep time. Through electroencephalogram (EEG) and electromyogram (EMG) recording, we demonstrated that luteolin increased non-rapid eye movement (NREM) sleep time and decreased wake time. To evaluate the underlying mechanism, we examined the effects of various pharmacological antagonists on the hypnotic effect of luteolin. The hypnotic effect of 3 mg/kg of luteolin was not affected by flumazenil, a GABAA receptorbenzodiazepine (GABAAR-BDZ) binding site antagonist, and bicuculine, a GABAAR-GABA binding site antagonist. On the other hand, the hypnotic effect of 3 mg/kg of luteolin was almost completely blocked by caffeine, an antagonist for both adenosine A1 and A2A receptor (A1R and A2AR), 8-Cyclopentyl-1,3-dipropylxanthine (DPCPX), an A1R antagonist, and SCH-58261, an A2AR antagonist. From the binding affinity assay, we have found that luteolin significantly binds to not only A1R but also A2AR with $IC_{50}$ of 1.19, $0.84{\mu}g/kg$, respectively. However, luteolin did not bind to either BDZ-receptor or GABAAR. From these results, it has been suggested that luteolin has hypnotic efficacy through A1R and A2AR binding.

Binding of $^3$H-Lectin from Kintoki Bean and Taro Tuber to Small Intestine of the Mouse (콩과 토란에서 추출한 $^3$H-Lectin의 마우스 소장에의 흡착량 정량)

  • 서영주
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.4
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    • pp.489-493
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    • 1993
  • The major objective of this study carried out was to compare the binding of Kintoki bean lectin (KBL) and Taro tuber lectin (TTL) to the mouse intestinal segments using $^3$H-labeled lectins and to assess the effect of such binding on the ability of the small intestine. Binding of $^3$H-KBL or $^3$H-TTL was studied under various conditions of time course, temperature, concentration, pH and additives of sugars, EDTA or unlabeled native lectin. The interaction of the lectins to intestinal tissue was stronger in KBL than in TTL, which was supposed to be the major reason for the stronger antinuritional enen of KBL. The optimal binding conditions were at 37$^{\circ}C$ for 60mins and at pH 7. The binding of both lectins were inhibited by fetuin and EDTA.

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Investigation of the Binding Site of CCR2 using 4-Azetidinyl-1-aryl-cyclohexane Derivatives: A Membrane Modeling and Molecular Dynamics Study

  • Kothandan, Gugan;Gadhe, Changdev G.;Cho, Seung Joo
    • Bulletin of the Korean Chemical Society
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    • v.34 no.11
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    • pp.3429-3443
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    • 2013
  • Chemokine receptor (CCR2) is a G protein-coupled receptor that contains seven transmembrane helices. Recent pharmaceutical research has focused on the antagonism of CCR2 and candidate drugs are currently undergoing clinical studies for the treatment of diseases like arthritis, multiple sclerosis, and type 2 diabetes. In this study, we analyzed the time dependent behavior of CCR2 docked with a potent 4-azetidinyl-1-aryl-cyclohexane (4AAC) derivative using molecular dynamics simulations (MDS) for 20 nanoseconds (ns). Homology modeling of CCR2 was performed and the 4AAC derivative was docked into this binding site. The docked model of selected conformations was then utilized to study the dynamic behavior of the 4AAC enzyme complexes inside lipid membrane. MDS of CCR2-16b of 4AAC complexes allowed us to refine the system since binding of an inhibitor to a receptor is a dynamic process and identify stable structures and better binding modes. Structure activity relationships (SAR) for 4AAC derivatives were investigated and reasons for the activities were determined. Probable binding pose for some CCR2 antagonists were determined from the perspectives of binding site. Initial modeling showed that Tyr49, Trp98, Ser101, Glu291, and additional residues are crucial for 4AAC binding, but MDS analysis showed that Ser101 may not be vital. 4AAC moved away from Ser101 and the hydrogen bonding between 4AAC and Ser101 vanished. The results of this study provide useful information regarding the structure-based drug design of CCR2 antagonists and additionally suggest key residues for further study by mutagenesis.