• Journal of the Korean Magnetic Resonance Society
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• v.22 no.1
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• pp.18-25
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• 2018

Replication protein A (RPA) is an essential single-stranded DNA binding protein in DNA processing. It is known that N terminal domain of RPA70 (RPA70N) recruits various protein partners including damage-response proteins such as p53, ATRIP, Rad9, and MRE11. Although the common binding residues of RPA70N were revealed, dynamic properties of the protein are not studied yet. In this study, we measured $^{15}N$ relaxation parameters ($T_1,\;T_2$ and heteronuclear NOE) of human RPA70N and analyzed them using model-free analysis. Our data showed that the two loops near the binding site experience fast time scale motion while the binding site does not. It suggests that the protein binding surface of RPA70N is mostly rigid for minimizing entropy cost of binding and the loops can experience conformational changes.

• Journal of Veterinary Clinics
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• v.15 no.2
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• pp.370-377
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• 1998

본 연구는 황색포도상구균의 병원성 인자인 alpha-toxin, casular polysaccharides (CPS)와 fibronectin-binding protein (FnBP)을 이용해 실험동물인 토끼에서의 항체가 형성 능과 공격접종 후 방어능에 관한 연구를 위하여 수행되었으며 향후 젖소 유방염 아단위 항원 을 이용한 백신개발 가능성을 탐색하고자 수행되어졌다. 황색포도상구균의 alpha-toxin, capsu18r polysaccharides (CPS)91 fibronectin-binding protein (FnBP) 항원을 이통재 효소면 역측정법을 통한 혈중 IgG 항체가수준을 측정하였으며 alpha-toxin, capsular polysaccharides (CPS)와 fibronectin-binding protein (FnBP)으로 면역시킨 토끼에서 대조군의 토끼보다 혈 중 항체가 수준이 1차 면역 이후 유의성 있게 높았다 (p<0.05). 백신에 사용된 alpha-toxin, capsular polysaccharides (CPS)와 fibronectin-binding protein (FnBP) 항원중 capsular polysaccharides (CPS)가 다른 alpha toxin과 fibronectin-binding protein (FnBP)의 혈중항 체가 수준과 비교하여 볼 때 비교적 낮은 수준이었다. 세균을 혈중으로 공격접종한 후 대조군 과 백신접종군의 혈중내 세균제거율에 있어 대조군에 비해 백신접종군에서 유의성 있게 낮은 균수를 보였다 (p<0.05). 또한 장기내 균수측정실험결과 대조군의 장기보다 백신접종군에서 유의성 있게 세균수가 낮게 출현하였다 (p<0.05). 결론적으로 본 연구결과 황색포도상구균의 3가지 항훤 alpha-toxin, capsular polysaccharide와 재조합 fibronectin binding protein을 이 퐁한 실험동물에서 안단위 유방염 백신은 방어능이 있다고 생각된다.

• Biomedical Science Letters
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• v.14 no.2
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• pp.77-81
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• 2008

Testis brain RNA-binding protein (TB-RBP) is a DNA/RNA binding protein. TB-RBP is mainly expressed in testis and brain and highly conserved protein with several functions, including chromosomal translocations, DNA repair, mitotic cell division, and mRNA transport, stabilization, and storage. In our previous study, we identified TB-RBP as an interacting partner for the catalytic subunit $(C{\alpha})$ of protein kinase A(PKA) and verified their interaction with several biochemical analyses. Here, we confirmed interaction between $C{\alpha}$. and TB-RBP in mammalian cells and determined the effect of $C{\alpha}$. on the function of TB-RBP. The activation of $C{\alpha}$. increased the TB-RBP function as a DNA-binding protein. These results suggest that the function of TB-RBP can be modulated by PKA and provide insights into the diverse role of PKA.

• Journal of Plant Biology
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• v.37 no.2
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• pp.151-158
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• 1994

Aleurone layers of normal and vp1 mutant maize kernels were extracted and centrifuged at 100,000g to yield a cytosol fraction. Binding of [3H]ABA cis, trans (＋)ABA to a soluble macromolecular components present in the cytosol was demonstrated by Sephadex chromatography and non-denaturing PAGE. The binding component was of high molecular weight and seems to be an aggregate of proteins. A rapid DEAE-cellulose filter method for assaying bound [3H]ABA to a soluble protein was adapted. Binding assays were performed with cytosol that had been preheated or incubated with several enzymes, indicating that heat and protease treatments disrupted the binding. This suggested that binding occurred to proteins. Some properties of the ABA binding proteins were described. The [3H]ABA binding were reduced dramatically when unlabeled ABA was added as a competitor, suggesting a specific binding of [3H]ABA. Gel filtration profiles and autoradiogram of [3H]ABA binding showed no difference in the binding components of Vp1 and vp1/vp1 mutant cytosol, indicating that Vp1 protein is not a sole ABA binding protein.

• The Korean Journal of Zoology
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• v.37 no.1
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• pp.40-48
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• 1994

세포분화에 따른 Small GTP-binding protein의 역할을 밝히기 위하여 Retinoic acid(RA)와 dibutyryl cyclic AMP(dbcAMP)로 분화를 유도한 F9 기형암종세포의 형태적인 변화와 함께 Small GTP-binding protein의 분포를 조사하였다. RA와 dbcAMP를 처리한 세포는 분화유도 5일경(초기 분화 단계)에 분명한 세포의 경계를 보이기 시작하여 7일경(분화 후기 단계)에는 거의 모든 세포가 등근 분화된 형태로 전환되었다. 이 분화과정 동안 세포막에는 많은 microvilli와 lamellopodia 같은 구조물이 나타났다. 아울러 초기 분화 단계에 많은 량의 laminin이 발현되었으며 분화 후기에 microtubule의 재분포가 관찰되었다. 세종류의 Small GTP-binding protein(25 23, 21 KD)이 F9 세포의 막성분과 세포질에서 관찰되었으며 분화가 진행됨에 따라서 세단백질 모두 증가되는 양상을 보였다 이러한 결과는 Small GTP-binding protein이 F9 세포의 분화에 특별한 기능을 가지고 있음을 시사해 주고 있다.

• Journal of Microbiology
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• v.44 no.5
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• pp.508-514
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• 2006

In this study, the double-stranded DNA-dependent activities of Deinococcus radiodurans RecA protein (Dr RecA) were characterized. The interactions of the Dr RecA protein with double-stranded DNA were determined, especially dsDNA-dependent ATP hydrolysis by the Dr RecA protein and the DNA strand exchange reaction, in which multiple branch points exist on a single RecA protein-DNA complex. A nucleotide cofactor (ATP or dATP ) was required for the Dr RecA protein binding to duplex DNA. In the presence of dATP, the nucleation step in the binding process occurred more rapidly than in the presence of ATP. Salts inhibited the binding of the Dr RecA protein to double-stranded DNA. Double-stranded DNA-dependent ATPase activities showed a different sensitivity to anion species. Glutamate had only a minimal effect on the double-stranded DNA-dependent ATPase activities, up to a concentration of 0.7 M. In the competition experiment for Dr RecA protein binding, the Dr RecA protein manifested a higher affinity to double-stranded DNA than was observed for single-stranded DNA.

• BMB Reports
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• v.38 no.1
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• pp.97-103
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• 2005

Under the condition of expression of $\lambda$ P protein at lethal level, the oriC DNA-binding activity is significantly affected in wild-type E. coli but not in the rpl mutant. In purified system, the $\lambda$ P protein inhibits the binding of both oriC DNA and ATP to the wild-type DnaA protein but not to the rpl DnaA protein. We conclude that the $\lambda$ P protein inhibits the binding of oriC DNA and ATP to the wild-type DnaA protein, which causes the inhibition of host DNA synthesis initiation that ultimately leads to bacterial death. A possible beneficial effect of this interaction of $\lambda$ P protein with E. coli DNA initiator protein DnaA for phage DNA replication has been proposed.

• YAKHAK HOEJI
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• v.42 no.1
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• pp.46-52
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• 1998

Retroviral integrase is required for integration of viral DNA into the host cell chromosome. Human immunodeficiency virus type-1 integrase was partially purified as a part of a fusion protein linked to a maltose-binding protein and characterized in terms of an endonucleolytic activity. The concentration of the fusion protein purified through an amylose column was about 12mg/ml. Indicating that the solubility of the fusion protein is highly increased by the presence of a maltose-binding protein, considering that the integrase protein alone is poorly solubilized. The endonucleolytic activity of the fusion protein was detected at 0.1 to 1.OmM $Mn^{++}$ ion, but not at any concentrations tested of $Mn^{++}$ ion.

• BMB Reports
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• v.43 no.6
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• pp.407-412
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• 2010

Homeodomain (HD) is a highly conserved DNA-binding domain composed of helix-turn-helix motif. Drosophila Vnd (Ventral nervous system defective) containing HD acts as a regulator to either enhance or suppress gene expression upon binding to its target sequence. In this study, kinetic analysis of Vnd binding to DNA was performed. The result demonstrates that DNA-binding affinity of the recombinant protein containing HD and NK2-specific domain (NK2-SD) was higher than that of the full-length Vnd. To access whether phosphorylation sites within HD and NK2-SD affect the interaction of the protein with the target sequence, alanine substitutions were introduced. The result shows that S631A mutation within NK2-SD does not contribute significantly to the DNA-binding affinity. However, S571A and T600A mutations within HD showed lower affinity for DNA binding. In addition, DNA-binding analysis using embryonic nuclear protein also demonstrates that Vnd interacts with other nuclear proteins, suggesting the existence of Vnd as a complex.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.242.3-243
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• 2003

The compound of 2"-O-benzoylcinnamaldehyde(CB-ph) is a derivative of 2"-hydroxycinnamaldehyde whcih is a methanol extract of cinnamomum cassia blume. It"s a new anti-cancer agent which has been showed to inhibit the growth of various tumor cells in vitro and in vivo. In order to investigate the effective drug concentration and bio-distribution of CB-ph, the plasma protein binding was studied. In this study, the degree of the binding of Cb-ph to various serum proteins, the binding parameters, the binding site of CB-ph in human serum albumin, and the effect of some extensive protein-binding drugs on the protein binding of CB-ph in human serum ablumin were investigated respectively by ultrafilteration and fluorescence spectrometry. (omitted)