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Kim, Cheorl-Ho,Lee, Tae-Kyun,Chung, Ji-Choen,Park, Won-Hwan,Kim, June-Ki,Kim, Yong-Ju,Park, Sun-Dong,Nam, Kyung-Soo,Kim, Yong-Sung 동국대학교 한의학연구소 1999 東國韓醫學硏究所論文集 Vol.7 No.2
FoLT-PCR 기술을 인체체질의하적 응용을 위하여 당뇨병연구에 사용하였다. 당뇨병은 제1형 및 제2형으로 나뉘는데 제1형은 인슈린비의존성(NIDDM)으로 당뇨병환자의 약60%이상을 차지하며, 제2형은 인슈린의존성(IDBM)으로 당뇨병환자의 30%미만을 차지한다. 이들은 대부분 후천적으로 환경중에서의 인슈린관련 유전자의 돌연변이에 의해 발병하는 것으로 알려져 있다. 이에, 본 연구에서는 당뇨병의 병인을 유전적변이와 체질의학적 관계에서 고찰하기 위하여 수행되었다. NIDDM환자와 IDDM환자를 대상으로 인슈린유전자를 증폭하여 제한효소절단 양상과 염기배열분석을 하였다. 비암호영역중 4개 위치 +216, +1045, +1367, 및 +1380에서 다형성을 보였으며 새로운 α4, α5, α6 및 β2이 α1과 β1가 이형(heterozygous)에서만 검출되며 α1은 우성이며 신규형들과 β1은 열성이었다. 이러한 당뇨병병인은 유전학적으로 체질의학과 깊은 관계를 가지는 것을 시사하였다. A simple and rapid FoLT(formamide low temperature)-PCR, whereby human genomic DNA from blood can be amplified without DNA preparative stps, is described using human insulin genes. By applicatin of FoLT-PCR in human insulin genes, intragenic polymorphism in non-coding regions of the human insulin gene was shown after amplification and analysis by restriction enzyme digestion. All nucleotide sequences were the same as the reported, and four necleotides, at 4 different positions were polymorphic, and polymorphic alleles α4, α5, α6, and β2 were identified. The new alleles were originated from homologous recombination between the α1 and β1 alleles, and the alleles were founded in heterozygotes only. Althouughallele α1 was dominant, the new alleles and β1 were recessive. From this results, it was suggested that the new method of FoLT-PCR was highly applicable in genetic variation analysis.
( Yong Tae Jeong ),( Ju Hye Yang ),( Xian Li ),( Geum Jin Kim ),( Dong Soo Kim ),( Cheorl Ho Kim ),( Min Kyun Na ),( Hyeun Wook Chang ) 영남대학교 약품개발연구소 2015 영남대학교 약품개발연구소 연구업적집 Vol.25 No.-
Hypoglycemic effects of ethylacetate extracts of Anguilla japonica (EMA) muscles in db/db mice were investigated. To understand the mechanism responsible for the hypoglycemic effects of EMA, the effects of EMA on AMP-activated protein kinase (AMPK) activation in L6 myotubes and in vivo using type II diabetic db/db mice were analyzed. In L6 myotubes, the phosphorylation degrees of AMPK and acetyl-CoA carboxylase (ACC) were markedly increased and glucose uptake was significantly (p<0.001) increased by EMA, compared with untretaed L6 myotubes. However, in L6 myotubes, these effects were abolished by compound C, an AMPK inhibitor. Moreover, EMA significantly reduced non-fasting blood glucose and serum insulin levels, and strongly induced AMPK phosphorylation in skeletal muscle tissues of db/db mice. EMA regulates glucose levels in L6 myotubes and in diabetic mice by activation of AMPK. Beneficial effects for diabetes treatment are indicated.
Molecular cloning and analysis of the <i>Thermus caldophilus</i> ADP-glucose pyrophosphorylase
Kim, Yong-Sam,Sohn, Hosung,Jin, Un-Ho,Suh, Seok-Jong,Lee, Sang Chul,Jeon, Jae Heung,Lee, Dae-Sil,Kim, Cheorl-Ho,Ko, Jeong Heon IPC Science and Technology Press 2007 Enzyme and microbial technology Vol.41 No.4
<P><B>Abstract</B></P><P>Previously, we have purified a highly thermostable ADP-glucose pyrophosphorylase (EC 2.7.7.27; AGPase) from <I>Thermus caldophilus</I> GK-24. In the present paper, we further report the molecular cloning and characterization of the thermostable bacterial AGPase. Using a 0.6-kb DNA probe obtained by polymerase chain reaction (PCR) with a primer set deduced from the <I>N</I>-terminal and internal amino acid sequence, the AGPase gene was cloned. The cloned AGPase gene had a 1245-bp open reading frame that encodes a protein of 414 amino acids. Then, the full-length AGPase gene was further cloned into the pHCE19T(II) vector and was expressed in <I>Escherichia coli</I> DH5α. Western analysis of the recombinant enzyme showed the same immunity as the wild-type enzyme with a molecular mass of ca. 46kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The kinetic parameters of the recombinant AGPase were basically similar to the wild type. The <I>N</I>-terminal region of <I>T. caldophilus</I> AGPase showed a partial similarity to that of <I>E. coli,</I> and R336 and P295 were identical to those of <I>E. coli</I> AGPase. In addition, sequence comparison revealed that R177 and P235 of <I>T. caldophilus</I> AGPase were aligned with K195 and K247 of <I>E. coli,</I> and K376 with R386, while K195 residue of <I>E. coli</I> was reported to be located in the glucose-1-phosphate (Glc-1-P) substrate binding region.</P>
Genetic Polymorphisms in Non-Coding Regions of the Human Insulin Gene
Kim, Yong-Sung,Kim, Jung-Ki,Kim, Cheorl-Ho 동국대학교 경주대학 1997 東國論集 Vol.16 No.1
Genetic polymorphism in non-coding regions of the human insulin gene was investigated. The 1.9 kb human insulin gene sequence, including the 5′to 3′flanking regions, was amplified by polymerase chain reaction, and analyzed by restriction enzyme analysis and direct DNA sequencing. The results showed that all nucleotide sequences in coding regions were the same as insulin gene sequences previously reported, and four necleotides, at positions +216, +1045, +1367, and +1380 in non-coding regions, were found to be polymorphic. In addition to previously identified polymorphic alleles α1(A-C-C-C) and β1(T-G-T-A), new nucleotide arrangements of α4(A-C-C-A), α5(A-G-C-C), α6(A-C-T-C), and β2(T-C-C-C) were identified. From the sequencing results, it was shown that these new alleles are derived from intragenic recombination within human insulin gene during chromosomal cross-over between the α1 and β1 alleles. The new alleles were detected only in heterozygous forms. Allele α1 was dominant from and the new variant alleles, as well as allele β1, appeared to be recessive in human insulin genes. The results suggested that intragenic recombination is possibly responsible for allelic divergence in the human insulin gene. Furthermore, this findings show the intragenic recombination to be a important factor for allelic divergence in higher eukaryotes and to be useful for future studies of human population genetics or insulin gene-related pathology. 인슈린의존성 당뇨병환자 2명과 인슈린비의존성 당뇨병환자 9명을 포함한 한국인24명을 대상으로 인슐린유전자의 유전적다형성을 해석하기 위하여 5′→3′ flanking영역을 포함하는 1.9kb유전자배열을 PCR법으로 증폭, 제한효소와 DNA염기배열결정을 통하여 분석하였다. 한국인 인슈린유전자에는 공통배열이 존재하였으며 비암호영역중 +216, +1045, +1367, 및 +1380위치에서 다형성을 나타내었다. 이미 보고된 다형성allele인 α1(A-C-C-C)과 β1(T-G-T-A) 이외에, 새로운 allele인 α4(A-C-C-A), α5(A-G-C-C), α6(A-C-T-C), 그리고 β2(T-C-C-C)가 신규로 확인되었다. DNA염기배열결과, 이러한 새로운 allele들의 생성은 α1과 β1 allele사이의 염색체전위과정중 내부재조합현상에 의한 것으로 추측되었으며 heterozygous형에서만 검출되었다. 그중 allele α1은 우성이며 새로운 변이allele들과 alleleβ1은 열성이었다. 본 결과들로부터 인체인슈린유전자를 비롯한 고등생물의 allele다양성은 내부재조합현상이 주요 요인임을 제시하였으며, 동시에 인체유전학과 당뇨병병리학 또는 체질의학적인 연구에 유용할 것을 시사하였다.
Kim, Yong-Sam,Son, Ok Lye,Lee, Ju Yeon,Kim, Sun Hee,Oh, Sejeong,Lee, Yoon Suk,Kim, Cheorl-Ho,Yoo, Jong Shin,Lee, Jeong-Hwa,Miyoshi, Eiji,Taniguchi, Naoyuki,Hanash, Samir M.,Yoo, Hyang Sook,Ko, Jeong H WILEY-VCH Verlag 2008 Proteomics Vol.8 No.16
<P>N-acetylglucosaminyltransferase V (GnT-V) has been reported to be upregulated in malignant cancer cells, and its targets have been sought after with regard to biomarker identification. The low capacity and high false positive rates of 2-DE gel-based lectin blots using phytohemagglutinin-L<SUB>4</SUB> (L-PHA) prompted us to develop a novel protocol for identifying GnT-V targets, in which serum proteins were subjected to immunodepletion, alkylation, and lectin precipitation using L-PHA coupled to avidin–agarose bead complexes, and tryptic digestion. Proteins captured by L-PHA conjugates were analyzed by a nano-LC-FT-ICR/LTQ MS. Here, we report 26 candidate biomarkers for colorectal cancer (CRC) that show 100% specificity and sensitivities of greater than 50%. Not only can these candidate proteins be used as analytes for validation, but the novel protocol described herein can be applied to biomarker discovery in nonCRCs. </P>
Kim, Yong-Sam,Kang, Hye-Yeon,Kim, Jin-Young,Oh, Sejeong,Kim, Cheorl-Ho,Ryu, Chun Jeih,Miyoshi, Eiji,Taniguchi, Naoyuki,Ko, Jeong Heon WILEY-VCH 2006 Proteomics Vol. No.
<P>To gain a better understanding of the mechanism underlying colon cancer and to search for potential markers of colon cancer prognosis, a comparative proteomic analysis of colon cancer WiDr cells was conducted using 2-DE and lectin blot, followed by identification based on ESI-MS. Through these approaches 14 proteins were identified as candidate target proteins for N-acetylglucosaminyl transferase V (GnT-V) that would be expected to be implicated in the progression of colon cancer. We selected protein tyrosine phosphatase kappa (PTPκ) as a model protein to validate this approach to the discovery of novel biomarkers in colon cancer. PTPκ underwent an aberrant glycosylation in GnT-V-overexpressing WiDr cells, and the aberrantly glycosylated PTPκ was vulnerable to proteolytic cleavage. The enhanced cleavage of PTPκ in GnT-V-overexpressing cells was responsible for the mitigation of the homophilic binding capacity, resulting in an increase in cancer cell migration.</P>
Shin, Deug-Yong,Kim, Cheorl-Ho,Kim, Kyoung-Sook,Lee, Young-Choon 東亞大學校附設遺傳工學硏究所 1998 遺傳工學硏究 Vol.- No.5
Two kinds of cDNA encoding mouse GalB1,3(4)GlcNAc α2,3-sialytransferase (mST3Gal III) and GalB1,4(3)GlcNAc α2,3-sialyltransferase (mST3Gal IV) were isolated from mouse brain cDNA library by means of a PCR-based approach. The cDNA sequences included an open reading frame coding for proteins of 374 and 333 amino acids. respectively, and the primary structure of these enzymes suggested a pultative domain structure consisting of four regions, like that in other glycosyltransferases. The deduced amino acid sequences of mST3Gal III and IV showed a 98% and 89% identity with rat ST3Gal III and human ST3Gal IV. respectively. Northern analysis indicated that the expression of mST3Gal III mRNA was abundant in heart. liver and adult brain, while that of mST3Gal IV mRNA was detected in all tissues tested except for testis. but the level was the highest in liver. Soluble forms of mST3Gal III and IV transiently expressed in COS cells exhibited enzyme activity toward acceptor substrates containing the terminal either GalB1.3GlcNAc or GalB1.4GlcNAc sequences. The substrate preferences of both enzymes were stronger for tetrasaccarides than for disaccarides.
Differential Distribution of Ganglioside GM3 in Seminiferous Tubule and Epididymis of Adult Rats
Jung, Kyu-Yong,Kim, Bo-Hyun,Cho, Mi-Ran,Kim, Hyoung-Min,Lee, Young-Choon,Kim, Cheorl-Ho,Kim, Jin-Kyeoung,Kim, Byung-Jin,Choo, Young-Kug The Pharmaceutical Society of Korea 2001 Archives of Pharmacal Research Vol.24 No.4
Gangliosides are ubiquitous membrane components in mammalian cells and are suggested to play important roles in various functions such as cell-cell interaction, adhesion, cell differentiation, growth control and signaling. Among all ganglio-series gangliosides, GM3 has the simplest carbohydrate structure, and has been shown as a major gangliosides, in male reproductive system. To study GM3 distribution in the seminiferous tubule and epididymis, frozen sections were stained with specific monoclonal antibody (MAb) against ganglioside GM3. In the seminiferous tubule of testis, pachytene spermatocytes and spermmatids expressed ganglioside GM3, but not in spermatogonia and sertoli cells. Spermatogonia and sertoli cells near the basement membrane were negatively reacted to anti-GM3. In the epididymis, GM3 was expressed only in some interstitial cells. Taken togethers, these results suggest that the expression of ganglioside GM3 in rat seminiferous tubule and epididymis is spatio-temporally regu lated during spermatogenesis.
( Min Kyun Na ),( Yong Tae Jeong ),( Xian Li ),( Fansi Jin ),( Seung Lark Hwang ),( Geum Jin Kim ),( Ju Hye Yang ),( Young Chae Chang ),( Dong Soo Kim ),( Cheorl Ho Kim ),( Hyeun Wook Chang ) 영남대학교 약품개발연구소 2015 영남대학교 약품개발연구소 연구업적집 Vol.25 No.-
The effect of butanol extracts of the skin of Anguilla japonica (BESA) on endoplasmic reticulum (ER) stress-induced insulin resistance in L6 myotubes was investigated. Western blotting revealed that BESA increased phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase, and stimulated glucose uptake in L6 myotubes. Stimulation of AMPK and glucose uptake by BESA were significantly (p<0.05) reduced by siRNA LKB1 or siRNA AMPK, compared with controls, suggesting that enhanced glucose uptake by BESA was predominantly accomplished via an LKB1-mediated AMPK activation pathway. In addition, BESA effectively reduced phosphorylation of ER stress markers, RNA-activated protein kinaselike ER resident kinase, JNK, and significantly (p<0.01) increased glucose uptake via AMPK activation in tunicamycin-treated L6 myotubes, compared with controls. Improvement of insulin sensitivity under ER stress conditions by BESA is predominantly accomplished via an LKB1- AMPK-dependent pathway.