• Bulletin of the Korean Chemical Society
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• 제32권9호
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• pp.3233-3238
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• 2011

The fluorescence of norfloxacin was quenched by various nucleotides. The ratio of the fluorescence intensities in the absence and presence of nucleotide was linearly dependent on nucleotide concentration, suggesting that quenching occurred through the formation of nonfluorescent norfloxacin-nucleotide complexes. The gradient of the linear relationship represented the equilibrium constant of complex formation; it decreased with increasing temperature. The slopes of van't Hoff plots constructed from the temperature-dependent equilibrium constants were positive in all cases, indicating that complex formation was energetically favorable - i.e., exothermic, with negative Gibb's free energy. The equilibrium constant increased when triphosphate was used instead of monophosphate. It also increased when the oxygen at the $C'_2$ position of the nucleotide was removed. Both enhancements were due to entropic effects: entropy decreased when complexes with AMP or GMP formed, while it increased when norfloxacin complexed with ATP, GTP, dAMP and dGMP.

• Journal of Applied Biological Chemistry
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• 제63권4호
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• pp.339-345
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• 2020

Euchrestaflavanone A (EFA) is a flavonoid found in the root bark of Cudrania tricuspidata. C. tricuspidata extract, widely used throughout Asia in traditional medicine, has been investigated phytochemically and biologically and is known to have anti-obesity, anti-inflammatory, and anti-tumor effects. It has been reported that C. tricuspidata extract also possesses anti-platelet effects; however, the mechanism of its anti-platelet and anti-thrombotic activities is yet to be elucidated. In this study, we investigated the effects of EFA on the modulation of platelet function using collagen-induced human platelets. Our results showed that EFA markedly inhibited platelet aggregation. Furthermore, it downregulated glycoprotein IIb/IIIa (αIIb/β3)-mediated signaling events, including platelet adhesion, granule secretion, thromboxane A2 production, and clot retraction, but upregulated the cyclic adenosine monophosphate-dependent pathway. Taken together, EFA possesses strong anti-platelet and anti-thrombotic properties and is a potential therapeutic drug candidate to prevent platelet-related thrombosis and cardiovascular disease.

• Archives of Pharmacal Research
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• 제21권6호
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• pp.712-717
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• 1998

The complete nucleotide sequence of the cDNA clone encoding rat skeletal muscle myosin- binding protein H (MyBP-H) was determined and amino acid sequence was deduced from the nucleotide sequence (GenBank accession number AF077338). The full-length cDNA of 1782 base pairs(bp) contains a single open reading frame of 1454 bp encoding a rat MyBP-H protein of the predicted molecular mass 52.7kDa and includes the common consensus 1CA__TG' protein binding motif. The cDNA sequence of rat MyBP-H show 92%, 84% and 41% homology with those of mouse, human and chicken, respectively. The protein contains tandem internal motifs array (-FN III-Ig C2-FN III- Ig C2-) in the C-terminal region which resembles to the immunoglobulin superfamily C2 and fibronectin type III motifs. The amino acid sequence of the C-terminal Ig C2 was highly conserved among MyBPs family and other thick filament binding proteins, suggesting that the C-terminal Ig C2 might play an important role in its function. All proteins belonging to MyBP-H member contains `RKPS` sequence which is assumed to be cAMP- and cGMP-dependent protein kinase A phosphorylation site. Computer analysis of the primary sequence of rat MyBP-H predicted 11 protein kinase C (PKC)phosphorylation site, 7 casein kinase II (CK2) phosphorylation site and 4N-myristoylation site.

• Journal of Microbiology and Biotechnology
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• 제15권3호
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• pp.573-578
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• 2005

Pyrimidine nucleotide N-ribosidase (pyrimidine 5'-nucleotide phosphoribo(deoxyribo)hydrolase/pyrimidine 5'-nucleotide nucleosidase, EC 3.2.2.10) catalyzes the breakdown of pyrimidine 5'-nucleotide into pyrimidine base and ribose(deoxyribo)-5-phosphate. However, detailed characteristics of the enzyme have not yet been reported. The enzyme was purified to homogeneity 327.9-fold with an overall yield of $6.1\%$ from Pseudomonas oleovorans ATCC 8062. The enzyme catalyzed cytidine monophosphate (CMP) and uridine monophosphate (UMP), but not adenosine monophosphate (AMP) and guanosine monophosphate (GMP). The enzyme optimally metabolized CMP at pH 6.0 and UMP at around 8.5, and the optimum temperature for the overall enzyme reaction was found to be $37^{\circ}C$. The $K_m$ values of the enzyme for CMP (at pH 6.0) and UMP (at pH 8.5) were 1.6 mM and 1.1 mM, respectively. AMP, deoxyCMP, and deoxyUMP were very effective inhibitors of the reaction. Double-reciprocal plots obtained in the absence and in the presence of AMP revealed that this inhibitory effect was of the mixed competitive type with respect to the breakdown of CMP and of the noncompetitive type with respect to the breakdown of UMP. In the presence of AMP, the enzyme followed sigmoid kinetics with respect to each substrate.

• 한국수산과학회지
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• 제17권5호
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• pp.368-372
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• 1984

보다 간편하게 핵산관련물질을 정량할 목적으로 고속액체크로마토그래피를 이용한 정량법을 검토하였으며, 이 방법을 이용하여 시판 수산건제품의 ATP분해생성물의 함량을 정량하였다. 장치는 HPLC/ALC-244(Waters Associates Inc.)를, 칼럼은 ${\mu}-Bondapak\;C_{18}$ stainless steel column($3.9mm\;i.d.{\times}30.0cm$)을, 검출기는 UV검출기(254nm)를, integrator는 Yanagimoto system-1000을 각각 사용하였으며, 이동상(移動相)으로서는 $1\%$ triethylamine${\codt}$phosphoric acid 혼액 (pH 6.5, $1\%$ triethylamine 용액을 phosphoric acid로써 pH조절)을 사용하고 2ml/min로 용리시키는 것이 가장 분리능이 좋았다. 용리되는 순서는 hypoxanthine, IMP, inosine, AMP, ADP 및 ATP의 순이었으며, retention time 20분이내에 분석이 완료되었다. 그러나 5'-IMP와 5'-GMP는 분리되지 않았다. 자건품(煮乾品)은 이노신산(IMP) 함량이 전반적으로 많았는데, 그 중에서도 벤뎅이가 가장 함량이 많았다. 소건품(素乾品)에는 inosine, hypoxanthine 함량이 많았으며, 조미건제품(調味乾製品)도 소건품과 비슷한 양상이었으나 IMP 함량이 비교적 높은 것은 조미할 때 핵산계조미료가 첨가되었기 때문이라고 볼 수 있었다.

• 한국의학물리학회지:의학물리
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• 제17권4호
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• pp.252-259
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• 2006

갭 정션(gap junction)은 다양한 세포에 분포되어 있으며 분자 수준의 작은 물질들이 자유롭게 교환되는 전기적 시냅스다. 망막에서, 갭 정션의 차단이 동물의 동작 감지(motion detection)에 실제적으로 어떤 영향을 주는지에 대해서는 거의 조사되지 않았다. 본 연구에서는, 망막 세포간의 전기적 시냅스를 조절하는 약물이 금붕어의 동작 감지에 어떠한 영향을 주는지를 조사하기 위해 시각운동 반응(optometer response, OMR)이 사용되었다. 갭 정션 차단제인 carbenoxolone, 8-bromo cyclic AMP, sodium nitroprusside (SNP), 8-bromo-cyclic GMP 등의 초자체 내 주사는 광- 및 암-상태에서 모두 OMR을 감소시켰다. 광-상태에서 dopamine, SKF-38393 및 eticlopride의 주사는 OMR을 감소시킨 반면 SCH-23390의 주사는 OMR을 증가시켰다. 암-상태에서는 결과가 반대로 나타났다: 즉 dopamine, SKF-38393 및 eticlopride의 주사는 OMR을 증가시킨 반면 SCH-23390의 주사는 OMR을 감소시켰다. 이러한 결과는 망막 세포들 사이의 갭 정션이 금붕어의 동작 감지에 중요한 역할을 담당하고 있음을 시사한다.

• 한국식품과학회지
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• 제15권3호
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• pp.245-251
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• 1983

쌀막걸리 담금중의 핵산분해효소들의 소장과 이효소들을 DEAE-cellulose column chromatography에 의해 분리정제하고 일반적인 성질을 검토함은 물론 핵산관련물질의 변화를 조사한 결과는 다음과 같았다. 1. 담금일수가 경과함에 따라 RNase는 담금 3 일까지 다소 증가하였다가 그후 점차 감소하였으며 PDase 및 PMase는 초기부터 활성이 감소하였다. 2. RNase의 최적pH는 5.0부근이었고, PDase, PMase는 6.0부근이었으며 pH안정성은 대체로 $pH6.0{\sim}7.0$부근이었다. 3. RNase 및 PDase의 최적온도는 $55{\sim}60^{\circ}C$이고, PMase는 50부근이었으며 열안정성은 RNase는 $100^{\circ}C$ PDase는 $80^{\circ}C$에서 10분간반응으로 각각 80%, 90%실활되었으며 PMase는 $70^{\circ}C$에서 10분간 반응으로 거의 샐활되었다. 4. RNase는 $CU^{++},\;Zn{++}$, PMase는 $10^{-3}M\;Na_{2}HPO_{4}$처리로서 약 30%의 효소활성이 저해되었다. 5. 술덧담금 4일째까지 IMP는 증가하였으며 UMP, GMP, AMP는 감소하는 경향을 보였다.

• BMB Reports
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• 제33권4호
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• pp.289-293
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• 2000

Annexin I is a 37 kDa member of the annexin family of calcium-dependent phospholipid binding proteins. Annexin I plays regulatory roles in various cellular processes including cell proliferation and differentiation. Recently we found that annexin I is a heat shock protein (HSP) and displays a chaperone-like function. In this paper we investigated the function of annexin I as an ATPase using 1 to 32 amino acids deleted annexin I (${\Delta}-annexin$ I). ${\Delta}-Annexin$ I hydrolyzed ATP as determined by thin layer chromatography. The ability of ATP hydrolysis was inhibited by ADP, GTP and GDP, but not by the AMP, GMP and cAMP. In view of the ATP hydrolyzing function of HSP, the results support the function of annexin I as a HSP.

• 한국미생물·생명공학회지
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• 제21권3호
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• pp.276-280
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• 1993

Cell lytic enzyme, 5'-phosphodiesterase, and AMP-deaminase were used to produce yeast extract as a natural seasoning from beer yeast cells. Prior to the addition of cell lytic enzyme, heat treatment was performed to increase the cell wall degradation` the optimum condition of the cell lytic enzyme was 50C at pH 7.0. The production yields by the enzymatic method and conventional autolysis method were 42% and 35%, respectively. The total quantity of 5'-nucleotides, GMP and IMP, produced by enzymatic method was increased by 45% than that by the conventional method. Futhermore, the operation time of enzymatic method was only 6.5 hrs, significantly reduced from 24 hrs of the conventional method.

• Journal of Ginseng Research
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• 제39권4호
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• pp.354-364
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• 2015

Background: Intracellular $Ca^{2+}$($[Ca^{2+}]_i$) is a platelet aggregation-inducing molecule. Therefore, understanding the inhibitory mechanism of $[Ca^{2+}]_i$mobilization is very important to evaluate the antiplatelet effect of a substance. This study was carried out to understand the $Ca^{2+}$-antagonistic effect of total saponin from Korean Red Ginseng (KRG-TS). Methods: We investigated the $Ca^{2+}$-antagonistic effect of KRG-TS on cyclic nucleotides-associated phosphorylation of inositol 1,4,5-trisphosphate receptor type I ($IP_3RI$) and cyclic adenosine monophosphate (cAMP)-dependent protein kinase (PKA) in thrombin (0.05 U/mL)-stimulated human platelet aggregation. Results: The inhibition of $[Ca^{2+}]_i$ mobilization by KRG-TS was increased by a PKA inhibitor (Rp-8-BrcAMPS), which was more stronger than the inhibition by a cyclic guanosine monophosphate (cGMP)- dependent protein kinase (PKG) inhibitor (Rp-8-Br-cGMPS). In addition, Rp-8-Br-cAMPS inhibited phosphorylation of PKA catalytic subunit (PKAc) ($Thr^{197}$) by KRG-TS. The phosphorylation of $IP_3RI$ ($Ser^{1756}$) by KRG-TS was very strongly inhibited by Rp-8-Br-cAMPS compared with that by Rp-8-BrcGMPS. These results suggest that the inhibitory effect of $[Ca^{2+}]_i$ mobilization by KRG-TS is more strongly dependent on a cAMP/PKA pathway than a cGMP/PKG pathway. KRG-TS also inhibited the release of adenosine triphosphate and serotonin. In addition, only G-Rg3 of protopanaxadiol in KRG-TS inhibited thrombin-induced platelet aggregation. Conclusion: These results strongly indicate that KRG-TS is a potent beneficial compound that inhibits $[Ca^{2+}]_i$ mobilization in thrombin-platelet interactions, which may result in the prevention of platelet aggregation-mediated thrombotic disease.