• 대한의생명과학회지
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• 제26권1호
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• pp.47-50
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• 2020

Recently, decades of robust researches on degenerative brain disorder have been highlighted on the interactive connection of gut and brain. In terms of inflammatory cytokine production, others have shown that Nucleotide-Binding Oligomerization Domain Containing 2 (NOD2) is involved with Leucine-Rich Repeat Kinase 2 (LRRK2). HEK293T cells were transiently co-transfected with Myc-tagged LRRK2 and Flag-tagged NOD2 and then followed by co-immunoprecipitation assay. In this study, we provide the novel finding of physical protein-protein interaction between NOD2 and LRRK2. G2019S variant has shown stronger interactions against NOD2 than those of wild type LRRK2. In an axis of NOD2-LRRK2 communication, it is believed to pave a new way in the understanding of the bidirectional molecular mechanism of brain disorder, including Parkinson's disease into gut inflammatory disease, including Crohn's disease.

• Asian-Australasian Journal of Animal Sciences
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• 제3권2호
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• pp.107-114
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• 1990

The purpose of this investigation was to study the hematological response of Saudi Arabian Baladi fowl to protein rearing regimens. Males and females were subjected to the following 4 protein rearing regimens: conventional, C; reverse protein, RP; 2 single-stage low protein, $SS_1$ and $SS_2$ using 15% and 12% CP diets, respectively. Regimen effect was highly significant ($$p{\leq_-}.01$$) on BW, PCY, TP and U-Ac and significant ($$p{\leq_-}.05$$) on TL. Serum chol levels were not affected by regimen. In general $SS_{2}$ birds showed the lowest values for all parameters studied, except for PCV. However, the differences were not significant in each case. Age and sex effects were highly significant ($$p{\leq_-}.01$$) for all parameters, however, the regimen X sex interaction was not significant except for PCV. Regimen X age interaction, on the other hand, was highly significant ($$p{\leq_-}.01$$) only for BW, TP and U-Ac concentrations. The data may suggest that low levels of protein in the rearing regimen is an important factor influencing levels of the blood parameters studied. The data also indicate a lack of clear relationship between hen-day egg production and the blood parameters studied.

• BMB Reports
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• 제45권3호
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• pp.183-188
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• 2012

Participates in actin remodeling through Rac and receptor endocytosis via Rab5. Here, we used yeast two-hybrid system with Eps8 as bait to screen a human brain cDNA library. ITSN2 was identified as the novel binding factor of Eps8. The interaction between ITSN2 and Eps8 was demonstrated by the in vivo co-immunoprecipitation and colocalization assays and the in vitro GST pull-down assays. Furthermore, we mapped the interaction domains to the region between amino acids 260-306 of Eps8 and the coiled-coil domain of ITSN2. In addition, protein stability assays and immunofluorescence analysis showed ITSN2 overexpression induced the degradation of Eps8 proteins, which was markedly alleviated with the lysosome inhibitor NH4Cl treatment. Taken together, our results suggested ITSN2 interacts with Eps8 and stimulates the degradation of Eps8 proteins.

• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1216-1221
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• 2006

A protein array was fabricated for a miniaturized immunoassay using microcontact printing ($\mu$CP). A polydimethylsiloxane (PDMS) stamp with a 5 $\mu$m$\times$5 /$\mu$m dimension was molded from a silicon master developed by photolithography. Under optimal fabrication conditions, including the baking, incubation, and exposure time, a silicon master was successfully fabricated with a definite aspect ratio. An antibody fragment was utilized as the ink for the $\mu$CP, and transferred to an Au substrate because of the Au-thiol (-SH) interaction. The immobilization and antibody-antigen interaction were investigated with fluorescence microscopy. When human serum albumin (HSA) was applied to the protein array fabricated with an antibody against HSA, the detection limit was 100 pg/ml of HSA when using a secondary antibody labeled with a fluorescence tag. The fabricated protein array maintained its activity for 14 days.

• BMB Reports
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• 제28권4호
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• pp.363-367
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• 1995

E. coli methionyl-tRNA synthetase is one of the class I tRNA synthetases. The Tryptophane residue at the position 461 located in the C-terminal domain of the enzyme is a key amino acid for the interaction with the anticodon of $tRNA^{Met}$. W461 was replaced with other amino acids to determine the chemical requirement for the interaction with the anticodon of $tRNA^{Met}$. Saturation mutagenesis at the position 461 generated a total of 12 substitution mutants of methionyl-tRNA synthetase. All the mutants showed the same in vivo stability as the wild-type enzyme, suggesting that the amino acid substitutions did not cause severe conformational change of the protein The mutants containing tyrosine, phenylalanine, histidine and cysteine substitutions showed in vivo activity while all the other mutants did not. The comparison of the in vitro aminoacylation activities of these mutants showed that aromatic ring structure, Van der Waals volume and hydrogen bond potential of the amino acid residue at the position 461 are the major determinants for the interaction with the anticodon of $tRNA^{Met}$.

• Journal of Microbiology and Biotechnology
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• 제17권2호
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• pp.359-363
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• 2007

Because of the importance of the type III protein-secretion system in bacteria-plant interaction, its function in bacterial pathogenesis of plants has been intensively studied. To identity bacterial proteins interacting with Xanthomonas hrp gene products that are involved in pathogenicity, we performed the glutathione-bead binding analysis of Xanthomonas lysates containing GST-tagged Hrp proteins. Analysis of glutathione-bead bound proteins by 1-DE and MALDI-TOF has demonstrated that Avr proteins, RecA, and several components of the type III secretion system interact with HrpB protein. This proteomic approach could provide a powerful tool in finding interaction partners of Hrp proteins whose roles in host-pathogen interaction need further studies.

• International Journal of Computer Science & Network Security
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• 제21권3호
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• pp.41-47
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• 2021

Detection of protein-protein interactions (PPIs) remains essential for the development of therapies against diseases. Experimental studies to detect PPI are longer and more expensive. Today, with the availability of PPI data, several computer models for predicting PPIs have been proposed. One of the big challenges in this task is feature extraction. The relevance of the information extracted by some extraction techniques remains limited. In this work, we first propose an extraction method based on correlation relationships between the physicochemical properties of amino acids. The proposed method uses a correlation matrix obtained from the hydrophobicity and hydrophilicity properties that it then integrates in the calculation of the bigram. Then, we use the SVM algorithm to detect the presence of an interaction between 2 given proteins. Experimental results show that the proposed method obtains better performances compared to the approaches in the literature. It obtains performances of 94.75% in accuracy, 95.12% in precision and 96% in sensitivity on human HPRD protein data.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.631-638
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• 2008

Tandem affinity purification (TAP) method combined with LC-MS/MS is the most accurate and reliable way to study the interaction of proteins or proteomics in a genome-wide scale. For the first time, we used a TAP-tag as a mutagenic tool to disrupt protein interactions at the specific site. Although lots of commonly used mutational tools exist to study functions of a gene, such as deletional mutations and site-directed mutagenesis, each method has its own demerit. To test the usefulness of a TAP-tag as a mutagenic tool, we applied a TAP-tag to RNA polymerase II, which is the key enzyme of gene expression and is controlled by hundreds of transcription factors even to transcribe a gene. Our experiment is based on the hypothesis that there will be interrupted interactions between Pol II and transcription factors owing to the TAP-tag attached at the C-terminus of each subunit of Pol II, and the abnormality caused by interrupted protein interactions can be observed by measuring a cell-cycle of each yeast strain. From ten different TAP-tagged strains, Rpb7- and Rpb12-TAP-tagged strains show severe defects in growth rate and morphology. Without a heterodimer of Rpb4/Rpb7, only the ten subunits Pol II can conduct transcription normally, and there is no previously known function of Rpb7. The observed defect of the Rpb7-TAP-tagged strain shows that Rpb7 forms a complex with other proteins or compounds and the interruption of the interaction can interfere with the normal cell cycle and morphology of the cell and nucleus. This is a novel attempt to use a TAP-tag as a proteomic tool to study protein interactions.

• Biomolecules & Therapeutics
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• 제8권3호
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• pp.276-280
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• 2000

In order to identify the domains of the G$_{\alpha}$16 subunit G protein that are responsible for its activation by the Interleukin-8 receptor, a serious of chimeras between G$_{\alpha}$16 and G$_{\alpha}$11 were assessed for their abilities to be activated by these receptors. Co-expression of IL-8 receptor and chimeras in which the carboxyl-terminal regions of G$_{\alpha}$11 were replaced from 30 up to 156 amino acid residues with the corresponding regions of G$_{\alpha}$16 demonstrated that C-terminal 156 amino acid residues of the G$_{\alpha}$16 were not sufficient to confer IL-8 receptor interaction specificity. Testing of a reciprocal serious of chimeras composed of G$_{\alpha}$16 sequences at the amino terminus and G$_{\alpha}$11 sequences at the carboxyl terminals revealed that sequences extending from the amino tar- minus to amino acid 209 of G$_{\alpha}$16 were sufficient to 7ndow the chimera with 75-80% of interaction specificity for 7-8-induced activation. These results suggest th,.7t combined interactions of the C-terminal 30 amino acid residues and certain domains extending from the arts.ino terminus to amino acid 209 of Gal 6 protein may be involved in its couplings to IL-8 receptor.tain domains extending from the arts.ino terminus to amino acid 209 of Gal 6 protein may be involved in its couplings to IL-8 receptor.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.639-642
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• 2003

본 연구는 후각 수용 단백질인 ODR10를 GST와 Histidine tag를 각각 N 말단과 C 말단에 삽입한 후 두 가지의 발현 벡터에 넣어 대장균에서 발현시켰다. 부분 정제된 단백질을 QCM의 수정진동자에 코팅한 후 여러 종류의 냄새 분자와의 상호 작용을 관찰하였다. 발현양은 적었지만 QCM실험 결과 발현된 단백질이 diacetyl과 반응한다는 것을 알 수 있었다. ODR10 단백질과 diacetyl의 결합 정도는 다른 냄새 분자와 비교했을 때 $5{\sim}10$배 정도 차이가 났으며 이를 통해 후각 수용 단백질을 발현시킨 대장균 세포들을 후각센서를 개발하는데 사용할 수 있다는 것을 알 수 있었다. 또한 현재까지는 1000가지 이상 존재한다고 알려진 후각 수용 단백질들이 어떤 냄새 분자와 특이적인 결합성을 가지는지 조사하기 위해서는 복잡하고 시간이 오래 걸리는 실험을 해야 했었지만, 대장균에서 발현시키는 시스템을 통해 경제적이고 효율적으로 조사를 할 수 있게 되었다.