• Interdisciplinary Bio Central
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• 제3권2호
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• pp.8.1-8.6
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• 2011

Background: Thanks to the development of the mathematical/statistical reverse engineering and the high-throughput measuring biotechnology, lots of biologically meaningful genegene interaction networks have been revealed. Steady-state analysis of these systems provides an important clue to understand and to predict the systematic behaviours of the biological system. However, modeling such a complex and large-scale system is one of the challenging difficulties in systems biology. Results: We introduce a new stochastic modeling approach that can describe gene regulatory mechanisms by dividing two (DNA and protein) layers. Simple queuing system is employed to explain the DNA layer and the protein layer is modeled using G-networks which enable us to account for the post-translational protein interactions. Our method is applied to a transcription repression system and an active protein degradation system. The steady-state results suggest that the active protein degradation system is more sensitive but the transcription repression system might be more reliable than the transcription repression system. Conclusions: Our two layer stochastic model successfully describes the long-run behaviour of gene regulatory networks which consist of various mRNA/protein processes. The analytic solution of the G-networks enables us to extend our model to a large-scale system. A more reliable modeling approach could be achieved by cooperating with a real experimental study in synthetic biology.

• 정보저장시스템학회논문집
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• 제9권1호
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• pp.28-31
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• 2013

We proposed a method to fabricate label-free protein sensor with sub-wavelength nanograting structures to be used for diagnosing acute myocardial infarction. A nickel stamp for the injection molding of nanograting integrated protein sensor was fabricated by electroforming process with high fidelity. By using metallic stamp, we replicated label-free protein sensor via injection molding, which is an outstanding method for low-cost and mass production of polymer products. Finally, we performed a feasibility test, examining cardiac troponin T (cTnT) and anti-cTnT interactions. From the results, we demonstrated that the fabricated protein sensor can provide information for the early and accurate detection of cardiac diseases such as acute myocardial infarction.

• Journal of Microbiology and Biotechnology
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• 제32권9호
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• pp.1209-1216
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• 2022

To better understand the effects of PEGylation and biotinylation on the delivery efficiency of proteins, the cationic protein lysozyme (LZ) and anionic protein bovine serum albumin (BSA) were chemically conjugated with poly(ethylene glycol) (PEG) and biotin-PEG to primary amine groups of proteins using N-hydroxysuccinimide reactions. Four types of protein conjugates were successfully prepared: PEGylated LZ (PEG-LZ), PEGylated BSA (PEG-BSA), biotin-PEG-conjugated LZ (Bio-PEG-LZ), and biotin-PEG-conjugated BSA (Bio-PEG-BSA). PEG-LZ and Bio-PEG-LZ exhibited a lower intracellular uptake than that of LZ in A549 human lung cancer cells (in a two-dimensional culture). However, Bio-PEG-BSA showed significantly improved intracellular delivery as compared to that of PEG-BSA and BSA, probably because of favorable interactions with cells via biotin receptors. For A549/fibroblast coculture spheroids, PEG-LZ and PEG-BSA exhibited significantly decreased tissue penetration as compared with that of unmodified proteins. However, Bio-PEG-BSA showed tissue penetration comparable to that of unmodified BSA. In addition, citraconlyated LZ (Cit-LZ) showed reduced spheroid penetration as compared to that of LZ, probably owing to a decrease in protein charge. Taken together, chemical conjugation of targeting ligands-PEG to anionic proteins could be a promising strategy to improve intracellular delivery and in vivo activity, whereas modifications of cationic proteins should be more delicately designed.

• 한국축산식품학회지
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• 제43권3호
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• pp.540-551
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• 2023

Milk protein concentrate (MPC) is widely used to enhance the stability and texture of fermented dairy products. However, most research has focused on yogurt products, and the effects of MPC on sour cream characteristics remain unknown. Therefore, we investigated the effects of different MPC levels (0%, 1%, 2%, and 3% w/w) on the rheological, physicochemical, microbiological, and aroma characteristics of sour creams in this study. We found that MPC supplementation stimulated the growth of lactic acid bacteria (LAB) in sour creams, resulting in higher acidity than that in the control sample due to the lactic acid produced by LAB. Three aroma compounds, acetaldehyde, diacetyl, and acetoin, were detected in all sour cream samples. All sour creams showed shear-thinning behavior (n=0.41-0.50), and the addition of MPC led to an increase in the rheological parameters (η_a,50, K, G', and G"). In particular, sour cream with 3% MPC showed the best elastic property owing to the interaction between denatured whey protein and caseins. In addition, these protein interactions resulted in the formation of a gel network, which enhanced the water-holding capacity and improved the whey separation. These findings revealed that MPC can be used as a supplementary protein to improve the rheological and physicochemical characteristics of sour cream.

• 생명과학회지
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• 제22권12호
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• pp.1608-1613
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• 2012

KIF5/Kinesin-I는 경쇄(light chain)를 통하여 결합함으로써 다양한 운반체들을 미세소관을 따라 운반한다. Kinesin light chains (KLCs)은 tetratricopeptide repeat (TPR) 영역을 매개로 운반체와 결합한다. 현재까지 KLCs와 결합하는 많은 운반체들이 확인되었으나 KLCs가 어떻게 특정운반체를 인식하여 결합하는지는 아직 확실히 밝혀지지 않았다. 본 연구에서 KLC1의 TPR 영역과 결합하는 단백질을 분리하기 위하여 효모 two-hybrid system을 이용하여 탐색한 결과 amyloid precursor protein (APP)과 결합하는 것으로 보고된 protein interacting with APP tail 1 (PAT1)을 분리하였다. KLC1은 PAT1의 C-말단 부위와 결합하며, PAT1은 KLC1의 TPR 영역을 포함한 부위와 결합함을 효모 two-hybrid assay로 확인하였다. 또한 PAT1는 KLC2와도 결합하였지만 kinesin heavy chains (KHCs)인 KIF5A, KIF5B, KIF5C와는 결합하지 않았다. 단백질간 결합은 glutathione S-transferase (GST) pull-down assay와 공동면역침강으로도 확인하였다. 생쥐의 뇌 파쇄액을 PAT1 항체와 APP 항체로 면역침강을 행한 결과 KLC와 KHCs가 같이 침강하였다. 이러한 결과들은 PAT1이 Kinesin-I와 APP 포함 소포간의 상호작용을 매개한다는 것을 시사한다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권5호
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• pp.455-461
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• 2006

In this study, we have described a method for the fabrication of a protein chip on silicon substrate using hydrophobic thin film and microfluidic channels, for the simultaneous detection of multiple targets in samples. The use of hydrophobic thin film provides for a physical, chemical, and biological barrier for protein patterning. The microfluidic channels create four protein patterned strips on the silicon surfaces with a high signal-to-noise ratio. The feasibility of the protein chips was determined in order to discriminate between each protein interaction in a mixture sample that included biotin, ovalbumin, hepatitis B antigen, and hepatitis C antigen. In the fabrication of the multiplexed assay system, the utilization of the hydrophobic thin film and the microfluidic networks constitutes a more convenient method for the development of biosensors or biochips. This technique may be applicable to the simultaneous evaluation of multiple protein-protein interactions.

• 한국정보과학회논문지:시스템및이론
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• 제33권5호
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• pp.267-281
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• 2006

단백질-단백질 상호작용(PPI : Protein-Protein Interaction) 데이타는 생물체가 어떠한 메커니즘으로 생명을 유지하는지에 대한 정보를 담고 있다. 질병 연구나 신약 연구를 위해서 PPI 데이타를 이용한 많은 연구들이 이루어지고 있다. 이러한 PPI 데이타의 크기는 Yeast-two-hybrid, Mass spectrometry과 Correlated mRNh expression과 같은 방법들로 인하여 점차 그 증가량이 커지고 있다. 따라서 단백질-단백질 상호작용 데이타의 방대한 양과 복잡한 구조로 인하여 사람이 직접 분석하는 것은 불가능하다. 다행히도 PPI 데이타는 단백질은 노드로, 상호작용은 에지로 표현함으로써 전산학의 그래프 구조로 추상화될 수 있다. 본 논문에서는 방대한 단백질-단백질 상호작용 데이타를 연구자가 다양한 방법으로 손쉽게 분석할 수 있는 워크벤치(workbench) 시스템인 Proteinca (PROTEin INteraction CAbaret)에 대하여 소개한다. Proteinca는 다앙한 데이타베이스의 PPI 데이타를 그래프이론 기반의 분석 기능들을 제공하며, 그래프로 가시화하여 사용자가 직관적으로 이해할 수 있도록 도와준다. 또한, 중력 모델 기반의 간략화 방법을 제공하여 사용자에게 중요 단백질 중심의 가시화를 제공한다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권3호
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• pp.1003-1008
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• 2016

Breast cancer is now the leading cause of cancer death in women worldwide. Cancer progression is driven not only by cancer cell intrinsic alterations and interactions with tumor microenvironment, but also by systemic effects. Integration of multiple profiling data may provide insights into the underlying molecular mechanisms of complex systemic processes. We performed a bioinformatic analysis of two public available microarray datasets for breast tumor stroma and peripheral blood mononuclear cells, featuring integrated transcriptomics data, protein-protein interactions (PPIs) and protein subcellular localization, to identify genes and biological pathways that contribute to dialogue between tumor stroma and the peripheral circulation. Genes of the integrin family as well as CXCR4 proved to be hub nodes of the crosstalk network and may play an important role in response to stroma-derived chemoattractants. This study pointed to potential for development of therapeutic strategies that target systemic signals travelling through the circulation and interdict tumor cell recruitment.

• Genomics & Informatics
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• 제12권2호
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• pp.64-70
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• 2014

Human papillomavirus (HPV) infection is the leading cause of cancer mortality among women worldwide. The life-threatening infection caused by HPV demands the need for designing anticancerous drugs. In the recent years, different compounds from natural origins, such as carrageenan, curcumin, epigallocatechin gallate, indole-3-carbinol, jaceosidin, and withaferin, have been used as a hopeful source of anticancer therapy. These compounds have been shown to suppress HPV infection by different researchers. In the present study, we explored these natural inhibitors against E6 oncoprotein of high-risk HPV-16, which is known to inactivate the p53 tumor suppressor protein. A robust homology model of HPV-16 E6 was built to anticipate the interaction mechanism of E6 oncoprotein with natural inhibitory molecules using a structure-based drug designing approach. Docking analysis showed the interaction of these natural compounds with the p53-binding site of E6 protein residues 113-122 (CQKPLCPEEK) and helped the restoration of p53 functioning. Docking analysis, besides helping in silico validation of natural compounds, also helps understand molecular mechanisms of protein-ligand interactions.

• Journal of Plant Biotechnology
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• 제49권4호
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• pp.300-306
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• 2022

The mitogen-activated protein kinase (MAPK) signaling cascade is essential for a wide range of cellular responses in plants, including defense responses, responses to abiotic stress, hormone signaling, and developmental processes. Recent investigations have shown that the stress, ethylene, and MAPK signaling pathways negatively affect the formation of nitrogen-fixing nodules by directly modulating the symbiotic signaling components. However, the molecular mechanisms underlying the defense responses mediated by MAPK signaling in the organogenesis of nitrogen-fixing nodules remain unclear. In the present study, I demonstrate that the Medicago truncatula mitogen-activated protein kinase kinase 5 (MtMKK5)-Medicago truncatula mitogen-activated protein kinase 3/6 (MtMPK3/6) signaling module, expressed specifically in the symbiotic nodules, promotes defense signaling, but not ethylene signaling pathways, thereby inhibiting nodule development in M. truncatula. U0126 treatment resulted in increased cell division in the nodule meristem zone due to the inhibition of MAPK signaling. The phosphorylated TEY motif in the activation domain of MtMPK3/6 was the target domain associated with specific interactions with MtMKK5. I have confirmed the physical interactions between M. truncatula nodule inception (MtNIN) and MtMPK3/6. In the presence of high expression levels of the defense-related genes FRK1 and WRKY29, MtMKK5a overexpression significantly enhanced the defense responses of Arabidopsis against Pseudomonas syringae pv. tomato DC3000 (Pst DC3000). Overall, my data show that the negative regulation of symbiotic nitrogen-fixing nodule organogenesis by defense signaling pathways is mediated by the MtMKK5-MtMPK3/6 module.