• Journal of Acupuncture Research
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• v.23 no.3
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• pp.177-190
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• 2006

Objectives : It has long been known about the anticancer effect of GRR-HAS, however, it has not been systemically determined the differentially regulated genes by GRR-HAS in cancer cells. The purpose of this study is to screen the GRR-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cell lines. Oligonucleotide microarray and proteomic approaches were employed to screen the differential expression genes. Methods : GRR~HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of GRR-HAS (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of GRR-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cells in all concentrations(0.1, 0.5, 1.5, 10,$20mg/m{\ell}$). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 320 with 6 up-regulated and 314 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. One down -regulated protein was protein disulfide isomerase and up-regulated proteins were fatty acid binding protein 1 and 14-3-3 gan1lTIa protein by $1.5mg/m{\ell}$ of CRR-HAS. Discussion : This study showed the comprehensive gene expression analysis using oligonucleotide microarray for the screening of GRR-HAS mediated differentially regulated genes. These results will provide a better application of GRR-HAS in cancer field and drug target development.

• BMB Reports
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• v.37 no.1
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• pp.107-113
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• 2004

Since the completion of the genome project of the nematode C. elegans in 1998, functional genomic approaches have been applied to elucidate the gene and protein networks in this model organism. The recent completion of the whole genome of C. briggsae, a close sister species of C. elegans, now makes it possible to employ the comparative genomic approaches for identifying regulatory mechanisms that are conserved in these species and to make more precise annotation of the predicted genes. RNA interference (RNAi) screenings in C. elegans have been performed to screen the whole genome for the genes whose mutations give rise to specific phenotypes of interest. RNAi screens can also be used to identify genes that act genetically together with a gene of interest. Microarray experiments have been very useful in identifying genes that exhibit co-regulated expression profiles in given genetic or environmental conditions. Proteomic approaches also can be applied to the nematode, just as in other species whose genomes are known. With all these functional genomic tools, genetics will still remain an important tool for gene function studies in the post genome era. New breakthroughs in C. elegans biology, such as establishing a feasible gene knockout method, immortalized cell lines, or identifying viruses that can be used as vectors for introducing exogenous gene constructs into the worms, will augment the usage of this small organism for genome-wide biology.

• BMB Reports
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• v.49 no.11
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• pp.607-611
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• 2016

The Cancer Genome Atlas (TCGA) has compiled genomic, epigenomic, and proteomic data from more than 10,000 samples derived from 33 types of cancer, aiming to improve our understanding of the molecular basis of cancer development. Availability of these genome-wide information provides an unprecedented opportunity for uncovering new key regulators of signaling pathways or new roles of pre-existing members in pathways. To take advantage of the advancement, it will be necessary to learn systematic approaches that can help to uncover novel genes reflecting genetic alterations, prognosis, or response to treatments. This minireview describes the updated status of TCGA project and explains how to use TCGA data.

• BMB Reports
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• v.41 no.3
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• pp.177-183
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• 2008

Ubiquitin is an essential, highly-conserved small regulatory protein in eukaryotic cells. It covalently modifies a wide variety of targeted proteins in the forms of monomer and polymers, altering the conformation and binding properties of the proteins and thus regulating proteasomal delivery, protein activities and localization. Mass spectrometry has emerged as an indispensable tool for in-depth characterization of protein ubiquitination. Ubiquitinated proteins in cell lysates are usually enriched by affinity chromatography and subsequently analyzed by mass spectrometry for identification and quantification. Ubiquitin-conjugated amino acid residues can be determined by unique mass shift caused by the modification. Moreover, the complex structure of polyubiquitin chains on substrates can be dissected by bottom-up and middle-down mass spectrometric approaches, revealing potential novel functions of polyubiquitin linkages. Here I review the advances and caveats of these strategies, emphasizing caution in the validation of ubiquitinated proteins and in the interpretation of raw data.

• BMB Reports
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• v.37 no.1
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• pp.35-44
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• 2004

Recently produced information on post-translational modifications makes it possible to interpret their biological regulation with new insights. Various protein modifications finely tune the cellular functions of each protein. Understanding the relationship between post-translational modifications and functional changes ("post-translatomics") is another enormous project, not unlike the human genome project. Proteomics, combined with separation technology and mass spectrometry, makes it possible to dissect and characterize the individual parts of post-translational modifications and provide a systemic analysis. Systemic analysis of post-translational modifications in various signaling pathways has been applied to illustrate the kinetics of modifications. Availability will advance new technologies that improve sensitivity and peptide coverage. The progress of "post-translatomics", novel analytical technologies that are rapidly emerging, offer a great potential for determining the details of the modification sites.

• Genomics & Informatics
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• v.14 no.1
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• pp.2-11
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• 2016

The advances in mass spectrometry-based proteomics technologies have enabled the generation of global proteome data from tissue or body fluid samples collected from a broad spectrum of human diseases. Comparative proteomic analysis of global proteome data identifies and prioritizes the proteins showing altered abundances, called differentially expressed proteins (DEPs), in disease samples, compared to control samples. Protein biomarker candidates that can serve as indicators of disease states are then selected as key molecules among these proteins. Recently, it has been addressed that cellular pathways can provide better indications of disease states than individual molecules and also network analysis of the DEPs enables effective identification of cellular pathways altered in disease conditions and key molecules representing the altered cellular pathways. Accordingly, a number of network-based approaches to identify disease-related pathways and representative molecules of such pathways have been developed. In this review, we summarize analytical platforms for network-based protein biomarker discovery and key components in the platforms.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.203-205
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• 2002

Light is an environmental signal that regulates photomovement and main energy source of photosynthesis in the cyanobacterium Synechocystis sp. PCC 6803 (Syn6803). Syn6803 is a popular model system for study of plant functional genomics. In this report, we adopted 2D gel based proteomics study to investigate proteins related with the light absorption and photo-protection in Syn6803. More than 700 proteins were detected on the SDS-gels stained with silver nitrate. Several proteins showing different expression level under various light conditions were identified with MALDI-TOF Mass spectrometry. As a comparison, we also conducted ICAT-based proteome study using WT and cphl (cyanobacterial phytochrome 1) mutant. A cphl deletion led to changes in the expression of proteins involved in translation, photosynthesis including photosystem and CO2 fixation, and cellular regulation. We are currently involved in TAP-tagging method to study protein-protein interactions in search for the molecular component involved in the light signal transduction of Syn6803 photomovement.

• Mass Spectrometry Letters
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• v.10 no.3
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• pp.71-78
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• 2019

For several decades, cancer has been the primary cause of mortality worldwide. New diagnosis and regimens have been developed to improve the chemotherapeutic efficacy and the quality of life of the patients. However, cancer tissues are complex and difficult to assess. Understanding the various properties of the tumor and its environment is crucial for cancer and pharmaceutical research. Several analytical techniques have been providing new insights into cancer research. Recently, matrix-assisted laser desorption ionization (MALDI)-mass spectrometry imaging (MSI), an advanced analytical technique, has been applied to translational research. Proteomic and lipidomic profiling obtained by MALDI-MSI has been critical for biomarker discovery and for monitoring heterogenous tumor tissues. In this review, we discuss technical approaches, benefits and recent applications of MALDI-MSI as a valuable tool in cancer research, namely for diagnosis, therapy, prognosis.

• Proceedings of the Korean Society of Life Science Conference
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• 2000.06a
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• pp.9-10
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• 2000

The aging process is multifactorial and results from the combined effects of inherited(genetic) and acquired factors including life style, food habits, physical activity, and diseases. That give rise to the various approaches in aging. We are trying to study biological changes with aging, In detail we are focused on gene and protein function accompanied by normal or abnormal aging process, especially our efforts are aimed at revealing the functional relationship of proteins in aging as a final product of gene. We expect that proteomic approach to the study of protein function involved in aging should give us variety of integrated data to understand biological changes of long lived lives, We have applied expression proteomics to rat liver bred in dietary restriction or in at libitum to elucidate the effects of food habit on aging. Expression proteomics shows us protein profile in a selected tissue or cells as a whole and gives us the information about protein expression level, posttranslational modification and degenerative modification of expressed proteins. Comparative analysis of young and old rat liver by two dimensional gels shows that gene expression of several proteins was down regulated in old rats and some protein expression level is increased with aging. Dietary restriction slows down these changes of gene expression and in some proteins there's no difference in protein expression level at same ages in comparison with rats bred in at libitum. About forty protein was identified by peptide mass fingerprint with MALDI-TOF and rest of the protein of interest is in the course of identification, Also we are trying to make mitochondrial and cytosolic proteom reference map. These suborganelle proteom map will gives us the information about low abundance proteins and cellular localization of proteins. Proteomics is a growing methodology to study biological system. High throughput qualitative and qualitative aspect of this approach will gives us large amount of integrated information and speed up our understanding about biological system

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.121-121
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• 2017

The productivity of rice has been influenced by various abiotic factors including temperature which cause to limitations to rice yield and quality. Rice yield and quality are adversely affected by high temperature globally. In the present study, four Korean four cultivars such as Dongan, Ilpum, Samkwang, Chucheong were investigated in order to explore molecular mechanisms of high temperature at seedling stage. Rice seedlings grown at $28/20^{\circ}C$ (day/night) were subjected to 7-day exposure to $38/28^{\circ}C$ for high-temperature stress, followed by 2-D based proteomic analysis on biological triplicates of each treatment. The growth characteristics demonstrated that Dongan is tolerant while Ilpum is sensitive to high-temperature stress. High temperature has an adverse effect in the seedling stage both in high temperature sensitive and tolerant cultivar. Two-dimensional gels stained with silver staining, a total of 722 differential expressed protein spots (${\geq}1.5-fold$) were identified using Progenesis SameSpot software. However, a total of 38 differentially expressed protein spots were analyzed by LTQ-FT-ICR MS. Of these, 9 proteins were significantly increased while 10 decreased under high-temperature treatment. Significant changes were associated with the proteins involved in the carbohydrate metabolism, photosynthesis, and stress responses. Proteome results revealed that high-temperature stress had an inhibitory effect on carbon fixation, ATP production, and photosynthetic machinery pathway. The expression level of mRNA is significantly correlated with the results obtained in the proteome investigation. Taken together, these findings provide a better understanding of the high-temperature resistance by proteomic approaches, providing valuable insight into improving the high-temperature stress tolerance in the global warming epoch.