• 생명과학회지
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• 제13권5호
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• pp.751-755
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• 2003

Cyclic AMP receptor proteins(CRP) activate many genes in Escherichia coli by binding of cAMP with not fully known mechanism. CRP existed as apo-CRP in the absence of cAMP, $CRP;(cAMP)_2$$_2$ at low(micromolar) cAMP concentration, or $CRP;(cAMP)_4$ at high(millimolar) concentration of cAMP. This study is designed to measure the thermal stability of S83G CRP, which substituted glycine for serine at amino acid 83 position, with CD spectrapolarimeter at 222nm by the constant elevation of temperature from $20^{\circ]C\; to\; 90^{\circ}C\; at\; 1^{\circ}C/min$. The non-linear regression analysis showed that melting temperatures were 68.4, 72.0, and $82.3^{\circ}C$ for no cAMP, 0.1mM cAMP, and 5mM cAMP, respectively. Result showed the strong thermal stability of CRP by binding of additional cAMP molecules to region between the hinge region and helix-turn-helix(HTH) motif at 5mM cAMP concentration.

• 생명과학회지
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• 제33권7호
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• pp.531-537
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• 2023

세포 내 수송 및 축삭 수송은 kinesin 및 cytoplasmic dynein과 같은 미세소관 의존성 모터단백질에 의해 운반된다. Kinesin은 미세소관을 따라 미세소관의 플러스 쪽 끝으로 이동하고, dynein은 미세소관의 마이너스 쪽 끝으로 이동한다. Kinesin-1은 kinesin superfamily protein (KIF)중에서 처음으로 확인된 kinesin으로, 카복실(C)-말단 영역과 cargo간 결합을 통해 세포내 소기관, 신경전달물질 수용체 및 mRNA-단백질 복합체를 포함한 다양한 cargo의 세포내 수송 기능을 수행한다. Kinesin-1은 다양한 cargo들을 수송하지만, kinsin-1과 cargo 사이를 매개하는 어댑터/스캐폴더 단백질은 아직 완전히 확인되지 않았다. KIF5A의 C-말단 영역과 상호 작용하는 어댑터 단백질을 규명하기 위해 효모 2-하이브리드 스크리닝을 하여, cyclin-dependent kinase 2-associated protein 1 (CDK2AP1)를 확인하였다. CDK2AP1은 KIF5A의 C-말단 영역에 결합하고 KIF3A, KIF5B, KIF5C 및 kinesin light chain 1 (KLC1)과는 결합하지 않았다. CDK2AP1의 C-말단 영역은 KIF5A와의 결합에 필수적이었다. HEK-293T 세포에 CDK2AP1 및 kinesin-1은 동시 발현하여 면역침강하면 CDK2AP1 및 kinesin-1은 같이 면역침강하였다. 그리고 CDK2AP1 및 kinesin-1은 세포내에서도 같은 위치에 발현하였다. 이러한 결과들은 KIF5A-CDK2AP1결합은 kinesin-1이 cargo를 운반할 때 kinesin-1과 cargo 사이를 연결하는 어댑터 단백질 역할을 시사한다.

• 한국식물병리학회지
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• 제11권1호
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• pp.73-79
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• 1995

Complementary DNAs (cDNAs) to the coat protein gene of an ordinary Korean strain of potato virus Y (PVY-OK) isolated from potato (cv. Superior) were synthesized and cloned into a plasmid pUC119 and sequenced. The RNA of the virus propagated in tobacco (Nicotinaa sylvestris) was extracted by the method of phenol extraction. The first strand of cDNAs to the coat protein penomic RNA of the virus was made by Moloney murine leukemia virus reverse transcriptase. The cDNA were synthesized and amplified by the method of polymerase chain reaction (PCR) using a pair of oligonucleotide primers. PVYCP3P and PVYCP3M. The size of cDNAs inserted in pUC119 plasmid was estimated as about 840 bp upon agarose gel electrophoresis. Double stranded cDNAs were transformed into the competent cell of E. coli JM109. Sequence analysis of cDNAs was conducted by the dideoxynucleotide chain termination method. Homology of cDNAs of the PVY-OK coat protein genomic RNA with those of PVY-O (Japan), PVY-T (Japan), PVY-TH (Japan), PVYN (The Netherlands),and PVYY (France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. Homology at the amino acid level turned out to the be 97.4%, 92.5%, 92.9%, 92.9% and 98.5%, respectively.

• Fisheries and Aquatic Sciences
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• 제2권2호
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• pp.161-166
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• 1999

The rigor-mortis progress of cultured olive flounder spiked at the brain started much faster than that of wild one. They attained full rigor state after 30 hrs at $0^{\circ}C$, 36 hrs at $5^{\circ}C$ and 50 hrs at $10^{\circ}C$ in the cultured flounder, while after 36 hrs at $0^{\circ}C$, 50 hrs at $5^{\circ}C$, and 60 hrs at $10^{\circ}C$ in the wild. ATP concentration in the muscle was around $5.9\mu mol/g$ for wild and $6.2\mu mol/g$ for cultured flounder. ATP breakdown progressed rapidly in $0^{\circ}C$ samples, followed by $5^{\circ}C$ and $10^{\circ}C$ samples. $Mg^{2+}$-ATPase activity of myofibrillar protein in the presence of 0.25mM CaCb was higher in cultured myofibri1lar protein than in wild one. $Mg^{2+}$-ATPase activities of myofibrillar protein increased during storage in samples stored at $0^{\circ}C$ and $5^{\circ}C$ while decreased in samples stored at $10^{\circ}C$. The level of breaking strength of muscle immediately after death was higher in the wild muscle than in the cultured muscle. The breaking strength reached maximum level at 10 hrs after death in both samples.

• 생명과학회지
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• 제24권1호
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• pp.14-19
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• 2014

Kinesin-1은 2개의 장쇄(KHCs, 또는 KIF5s)와 2개의 단쇄(KLCs)가 결합한 복합체로 되어 있다. 본 연구에서 효모 two-hybrid system을 이용하여 중추신경계의 신경세포에서 주로 발현되는 KIF5A와 결합하는 단백질을 탐색한 결과 phosphatidylinositol-3,5-bisphosphate ($PI(3,5)P_2$)의 5번 위치 인산을 제거하는 탈인산화효소 Fig4(Sac3)를 분리하였다. KIF5A는 Fig4의 C-말단과 결합함을 효모 two-hybrid assay로 확인하였다. Fig4는 KIF5A의 C-말단과 결합하지만, 두 개의 다른 장쇄인 KIF5B와 KIF5C 그리고 KLC1와는 결합하지 않았다. 단백질 간 결합을 glutathione S-transferase pull-down assay와 공동면역침강으로 추가 검증하였다. 생쥐의 뇌 파쇄액을 KIF5A 항체로 면역 침강한 결과 Fig4가 같이 침강하였다. 이러한 결과들은 kinesin-1이 Fig4와 결합한 단백질 복합체 혹은 운반체를 세포 내에서 운반함을 시사한다.

• Journal of Periodontal and Implant Science
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• 제39권sup2호
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• pp.279-286
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• 2009

Purpose: Gene therapy (ex vivo) has recently been used as a means of delivering bone morphogenetic proteins (BMPs) to sites of tissue regeneration. In the present study, we investigated the effect of co-transduction of adenoviruses expressing BMP-2 and BMP-7 on osteogenesisof C2C12 cells in vitro. Methods: A replication-defective human adenovirus 5 (Ad5) containing a cDNA for BMPs in the E1 region of the virus (Ad5BMP-2 and Ad5BMP-7) was constructed by in vivo homologous recombination. Functional activity of Ad5BMP-2 and Ad5BMP-7 were evaluated in mouse stromal cells (W20-17cells). C2C12 cells are transduced with various MOI (multiplicity of infection) of Ad5BMP-2 and Ad5BMP-7 to assess most effective and stable titer. Based on this result, C2C12 cells were transduced with Ad5BMP-2 and Ad5BMP-7 alone or by combination. BMPs expression, alkaline phosphatase (ALPase) activity, cell proliferation, and mineralization were assessed. Results: Ad5BMP-2 and Ad5BMP-7 are successfully transduced to W20-17 cells, and secreted BMPs stimulated cell differentiation. Also, C2C12 cells transduced with Ad5BMPs showed expression of BMPs and increased ALPaseactivity. In all groups, cell proliferation was observed over times. At 7days, cells co-transduced with Ad5BMP-2 and Ad5BMP-7 showed lower proliferation than the others. C2C12 cells co-transduced with Ad5BMP-2 and Ad5BMP-7 had greater ALPaseactivity than that would be predicted if effect of individual Ad5BMPs were additive. Little mineralized nodule formation was detected in cells transduced with individual Ad5BMPs. In contrast, Ad5BMP-2 and Ad5BMP-7 combination stimulated mineralization after culturing for 10 days in mineralizing medium. Conclusions: Present study demonstrated that adenoviruses expressing BMPs gene successfully produced BMPs protein and these BMPs stimulated cells to be differentiated into osteoblastic cells. In addition, the osteogenic activity of Ad5BMPs can be synergistically increased by co-transduction of cells with Ad5BMP-2 and Ad5BMP-7.

• 수산해양교육연구
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• 제23권3호
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• pp.461-467
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• 2011

This experiment was conducted to determine the each level of a newly developed squid liver powder (SLP) as a dietary protein source replacing fish meal (FM) in juvenile flounder. A diet excluding SLP (FM diet) and a commercial diet were two control diets. Six experimental diets were formulated to replace FM with three types of SLP at 5 and 10% level, respectively (SLP-A5, SLP-A10, SLP-B5, SLP-B10, SLP-C5, SLP-C10). Prior to the start of a feeding trial, fish were fed the commercial diet for 1 week to adjust to the experimental conditions. Fish with average weight of $22.8{\pm}0.40g$ ($mean{\pm}SD$), were randomly distributed to each aquarium as groups of 20 fish and fed the experimental diets in triplicate at satiation for 8 weeks. After 8 weeks, weight gain and specific growth rate of fish fed SLP-C10 was lower than those of fish fed FM diet and SLP-B5. Meanwhile, no significant difference was observed in WG and SGR among fish fed diets excluding SLP-C10. Feed efficiency (FE) and protein efficiency ratio (PER) of fish fed each level of three types of SLP were not different from those of fish fed FM diet. However, fish fed SLP-C5 and SLP-C10 has lower FE and PER than those of fish fed a commercial diet, SLP-A5 and SLP-B5. Therefore, these results indicate clearly that dietary SLP-B could be a good protein source to develop inexpensive commercial diet in juvenile flounder.

• 한국식품영양과학회지
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• 제18권2호
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• pp.195-198
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• 1989

도축장에서는 폐기되는 돼지 혈액으로부터 혈장을 분리하여, pH, 온도, 단백질함량에 따른 혈장 단백질 보수력의 변화를 조사하였다. 돼지 혈액으로부터 혈장분리는 1400g-force에서 가장 좋았다. 단백질 농도가 5%인 혈장액을 $85^{\circ}C$에서 30분간 가열하였을 때 pH가 증가함에 따라 혈장단백질보수력은 급격히 증가하다가 pH 7 이후에는 완만한 증가를 나타냈다. 단백질 농도가 5%이고 pH 7인 혈장액에서의 보수력은 gel화 온도가 높을수록 짧은 시간 내에 높은 보수력을 나타냈으며 가열 초기에 급격히 증가하다가 일정 가열 시간 이후에는 큰 변화를 나타내지 않았다. pH가 7인 혈장액을 $85^{\circ}C$에서 30분간 가열하였을 때 혈장 단백질의 농도가 증가함에 따라 단위 단백질 무게당 보수력은 감소하였다.

• The Korean Journal of Physiology and Pharmacology
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• 제16권3호
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• pp.211-217
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• 2012

Recent studies have demonstrated that nitric oxide (NO) activates transient receptor potential vanilloid subtype 1 (TRPV1) via S-nitrosylation of the channel protein. NO also modulates various cellular functions via activation of the soluble guanylyl cyclase (sGC)/protein kinase G (PKG) pathway and the direct modification of proteins. Thus, in the present study, we investigated whether NO could indirectly modulate the activity of TRPV1 via a cGMP/PKG-dependent pathway in cultured rat dorsal root ganglion (DRG) neurons. NO donors, sodium nitroprusside (SNP) and S-nitro-N-acetylpenicillamine (SNAP), decreased capsaicin-evoked currents ($I_{cap}$). NO scavengers, hemoglobin and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (CPTIO), prevented the inhibitory effect of SNP on $I_{cap}$. Membrane-permeable cGMP analogs, 8-bromoguanosine 3', 5'-cyclic monophosphate (8bromo-cGMP) and 8-(4chlorophenylthio)-guanosine 3',5'-cyclic monophosphate (8-pCPT-cGMP), and the guanylyl cyclase stimulator YC-1 mimicked the effect of SNP on $I_{cap}$. The PKG inhibitor KT5823 prevented the inhibition of $I_{cap}$ by SNP. These results suggest that NO can downregulate the function of TRPV1 through activation of the cGMP/PKG pathway in peripheral sensory neurons.

• 생물정신의학
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• 제5권2호
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• pp.227-234
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• 1998

Objective : Many evidences suggest that patients with bipolar disorder have functional abnormalities in their postreceptor signal transduction pathways, and mood stabilizing effect of lithium is exerted by modulating this dysfunctioning system. Carbamazepine, an antiepileptic agent, is also known to be effective in the treatment and prevention of bipolar disorder. But the precise mechanism of action of the drug is still poorly understood. This study was performed to elucidate the possible therapeutic mechanism of carbamazepine. Method : The effects of chronic carbamazepine administration on protein kinase A and protein kinase C activities in frontal cortex of rat brain after 2 weeks of drug administration were measured and compared with those of control subjects. Results : Mean(${\pm}SE$) value of activity(phosphate transfer ${\mu}mol/mg$ of $protein{\cdot}min$) of protein kinase A in control and test group was $0.249563{\pm}0.036$ and $0.539853{\pm}0.078$, and that of protein kinase C was $0.654817{\pm}0.053$ and $1.146205{\pm}0.052$ respectively, being increased in test group. And differences between the two groups were statistically significant for both enzymes(protein kinase A ; p<0.01, protein kinase C ; p<0.001). Conclusion : These results show that chronic carbamazepine administration increases protein kinase A and C activities, and concerning the possible mode of therapeutic action in bipolar disorder it is suggested that enhanced enzymes phosphorylate receptor-G-protein-effector complexes to dampen hyperfunctioning neuronal activity and thus stabilize the system.