• Journal of Microbiology and Biotechnology
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• 제18권5호
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• pp.968-974
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• 2008

Proteomic analysis led to identification of the proteins of Vibrio vulnificus that were induced upon exposure to INT-407 cells, and 7 of which belong to the functional categories such as amino acid transport/metabolism, nucleotide transport/metabolism, posttranslational modification/protein turnover/chaperones, and translation. Among the genes encoding the host-induced proteins, disruption of purH, trpD, tsaA, and groEL2 resulted in reduced cytotoxicity. The purH, trpD, and tsuA mutants showed impaired growth in the INT-407 lysate; however, the growth rate of the groEL2 mutant was not significantly changed, indicating that the possible roles of the host-induced proteins in the virulence of V. vulnificus are rather versatile.

• 생명과학회지
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• 제34권5호
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• pp.333-338
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• 2024

키네신-1은 모터 도메인이 있는 두 개의 중쇄(KHC 또는 KIF5)와 모터 도메인이 없는 두 개의 경쇄(KLC)로 구성된 이형사량체 단백질이다. KIF5에는 KIF5A, KIF5B 및 KIF5C의 세 가지 subtype이 있으며, 카르복실(C)-말단 영역을 제외하고는 아미노산 상동성이 높다. KIF5A는 세포 내에서 화물을 운반하며, KIF5A의 C-말단 영역에 결합하는 매개 단백질은 키네신-1과 화물 사이를 연결한다. 키네신-1의 세포내 수송을 조절하는 단백질은 아직 충분히 확인되지 않았다. 본 연구는 리소좀의 세포 내 수송에 관여하는 ADP-ribosylation GTPase-activating protein 1 (ArfGAP1)과 KIF5A와의 결합을 확인하였다. KIF5A는 ArfGAP1의 C-말단 영역에 결합하고, ArfGAP1은 KIF5A의 C-말단 영역에 결합하지만 KIF5B, KIF5C, 키네신 경쇄 1 (KLC1) 또는 KIF3A와는 결합하지 않았다. ArfGAP 도메인을 가진 다른 동질형인 SMAP1과는 결합하지 않았다. 세포에서 KIF5A는 ArfGAP1과 같은 위치에서 발현하며, KIF5A, KIF5B 및 KLC1와 같이 면역 침전하였다. 그러나, KIF3B와는 같이 면역 침전하지 않았다. 이러한 결과들은 키네신-1은 세포내 화물 수송에서 ArfGAP1에 의해 조절될 수 있음을 시사한다.

• Korean Journal of Acupuncture
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• 제29권2호
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• pp.260-270
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• 2012

Objectives : Hippocampus, a region of temporal lobe, plays an important role in the pathogenic mechanisms of brain diseases such as Alzheimer's disease, depression and temporal lobe epilepsy. This research is designed to investigate hippocampal changes after acupuncture stimulation at Shinmun(HT7) using 2-dimensional gel electrophoresis(2-DE). Methods : On postnatal-day 15, rat pups were randomly devided into Normal(NOR) or HT7 group. All of Pups kept with their mothers for 7 days, but pups in HT7 group received acupuncture stimulation at HT7 daily. On postnatal-day 21, hippocampus of each rat pup was dissceted 30 minutes after last acupuncture stimulation and the protein expressions were investigated using 2-DE. Results : After acupuncture stimulation at HT7, expression of 20 proteins were significantly increased. Succinate semialdehyde dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase-like, transketolase, aconitate hydratase and phosphoglucomutase-1 were related to glucose methabolism. Eukaryotic initiation factor(eIF) 4A-II, eIF 4A-III, mitochondrial Tu translation elongation factor and chain A of crystal structure of the 70-Kda heat shock cognate protein involve in the protein synthesis in ribosome. Tubulin ${\beta}$-4 chain, tubulin T ${\beta}$-15 and tubulin ${\alpha}$-1B chain comprise cytoskeleton. Glutathione S-transferase(GST) ${\omega}$-1, GST P and GST Yb-3 can reduce oxidative stress. ${\beta}$-soluble N-ethylmaleimide-sensitive fusion protein attachment protein is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus, glycerol-3-phosphate dehydrogenase plays a major role in lipid biosynthesis, creatine kinase U-type catalyses the conversion of creatine and consumes adenosine triphosphate to create phosphocreatine and adenosine diphosphate. Platelet-activating factor acetylhydrolase IB subunit alpha and voltage depedent anion-selective channel protein 2 were also increased. Conclusions : The results suggest that acupuncture stimulation at HT7 may enhance glucose and lipid metabolism, protein synthesis, cytoskeletal substance and anti-oxidative stress in hippocampus.

• Asian Pacific Journal of Cancer Prevention
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• 제15권8호
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• pp.3471-3476
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• 2014

Background: Aberrant expression of the microRNA-29 family is associated with tumorigenesis and cancer progression. As transport carriers, tumor-derived exosomes are released into the extracellular space and regulate multiple functions of target cells. Thus, we assessed the possibility that exosomes could transport microRNA-29c as a carrier and correlations between microRNA-29c and apoptosis of bladder cancer cells. Materials and Methods: A total of 28 cancer and adjacent tissues were examined by immunohistochemistry to detect BCL-2 and MCL-1 expression. Disease was Ta-T1 in 12 patients, T2-T4 in 16, grade 1 in 8, 2 in 8 and 3 in 12. The expression of microRNA-29c in cancer tissues was detected by quantitative reverse transcriptase PCR (QRT-PCR). An adenovirus containing microRNA-29c was used to infect the BIU-87 human bladder cancer cell line. MicroRNA-29c in exosomes was measured by QRT-PCR. After BIU-87 cells were induced by exosomes-derived microRNA-29c, QRT-PCR was used to detect the level of microRNA-29c. Apoptosis was examined by flow cytometry and BCL-2 and MCL-1 mRNA expressions were assessed by reverse transcription-polymerase chain reaction. Western blotting was used to determine the protein expression of BCL-2 and MCL-1. Results: The expressions of BCL-2 and MCL-1 protein were remarkably increased in bladder carcinoma (p<0.05), but was found mainly in the basal and suprabasal layers in adjacent tissues. The expression of microRNA-29c in cancer tissues was negatively correlated with the BCL-2 and MCL-1. The expression level of microRNA-29c in exosomes and BIU-87 cells from the experiment group was higher than that in control groups (p<0.05). Exosome-derived microRNA-29c induced apoptosis (p<0.01). Although only BCL-2 was reduced at the mRNA level, both BCL-2 and MCL-1 were reduced at the protein level. Conclusions: Human bladder cancer cells infected by microRNA-29c adenovirus can transport microRNA-29c via exosomes. Moreover, exosome-derived microRNA29c induces apoptosis in bladder cancer cells by down-regulating BCL-2 and MCL-1.

• The Korean Journal of Physiology and Pharmacology
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• 제4권2호
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• pp.91-97
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• 2000

The purpose of the present study was to determine the preventive effects of combined interventional trial of fish oil treatment and exercise training on insulin resistance of skeletal muscle in high-fat fed rats. Male Wistar rats were randomly divided into chow diet (CD), high-fat diet (HF), high-fat diet with fish oil (FO), high-fat diet with exercise training (EX), and FO+EX groups. The rats in control group were fed chow diet containing, as percents of calories, 58.9% carbohydrate, 12.4% fat, and 28.7% protein. High-fat diet provided 32% energy as lard, 18% as corn oil, 27% as carbohydrate and 23% as casein. The fish oil diet had the same composition as the high fat diet except that 100 g menhaden oil was substituted for corn oil. Insulin sensitivity was assessed by in vitro glucose transport in the soleus muscle after diet treatment and treadmill running for 4 weeks. While the FO or EX only partially prevented insulin resistance on glucose transport and visceral obesity induced by high-fat diet, these interventions completely corrected hyperinsulinemia and hyperglycemia from the high-fat diet. The rats in the FO+EX showed normalized insulin action on glucose transport, plasma chemicals and visceral fat mass. Insulin-mediated glucose transport was negatively associated with total visceral fat mass (r=－0.734; p＜0.000), plasma triglyceride (r=－0.403; p＜0.05) and lepin (r=－0.583; p＜0.001) concentrations with significance. Multiple stepwise regression analysis showed that only total visceral fat mass was independently associated with insulin-mediated glucose transport (r=－0.668; p＜0.000). In conclusion, combined interventional trial of FO+EX recovered insulin resistance on glucose transport of skeletal muscle induced by high-fat diet. Visceral fat mass might be more important factor than plasma TG and leptin to induce insulin resistance on glucose transport of skeletal muscle in high-fat fed rats.

• KSBB Journal
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• 제24권3호
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• pp.259-266
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• 2009

AfsR2 과발현 S. lividans TK21을 이용하여 DNA microarray를 수행하였다. 그 결과, phosphate starvation과 관련 있는 42개의 유전자들이 up-regulated 되었으며, 특히 SCO4139 (pstB, phosphate ABC transport system ATP-binding protein)와 SCO4142 (pstS, phosphate-binding protein precursor)는 afsR2가 phosphate와 같은 nutrient starvation에 적극적으로 관여한다는 것을 나타내며, SCO4228 (putative phosphate transport system regulatory protein)은 기존에 수행되었던 2D-electrophoresis 연구나 afsS null S. coelicolor를 이용한 DNA microarray 연구에서도 공통적으로 보고되었던 유전자로서 phosphate lilitation에 대한 afsR2의 효과가 지속적으로 검증되고 있음을 뜻한다. 또한 afsR2 과발현을 통해서 sigma factor인 SCO2954 (sigL)과 SCO5147 (sigE)의 발현이 유도되었으며 두 유전자의 구조적인 특징을 고려해 보았을 때 afsR2가 RNA polymerase와의 linker로서의 역할을 추측해 볼 수 있다. 뿐만 아니라 whi 관련 유전자들의 발현 또한 afsR2에 의해 증가되었다. 이는 afsR2가 단순히 2차 대사물질 생합성 조절에만 관여하는 것이 아니라 형태적 분화에 작용함으로써 최종적으로 여러 2차 대사물질의 합성을 유도한다고 말할 수 있다. 이러한 결과들을 토대로 afsR2가 기존에 항생제 생합성에만 관여하는 global regulatory 조절인자가 아닌 방선균이 stationary phase로 전환되는 시점에서 형태적 분화에 영향을 미치고 phosphate limitation stress를 줄여주는 2차 대사의 key-factor regulatory 유전자임을 알 수 있다.

• Journal of Animal Science and Technology
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• 제47권6호
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• pp.1087-1094
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• 2005

세포 생장과 대사에 있어서 glucose의 세포내 도입은 대부분 동물세포에 필수적이며 이와 같은 도입은 glucose transport protein에 의하여 수행된다. glucose transport protein 중에 GLUT4는 사람과 설치류의 지방조직과 골격근에 있어서 인슐린 자극에 의하여 glucose을 세포내로 도입하는 중요한 역할을 수행한다. 본 연구에서는 한우로부터 이와 같은 GLUT4 유전자를 동정하고 그 발현을 조사하였다. 한우 GLUT4 유전자는 1527bp의 open reading frame으로 구성되어 있으며 509개의 아미노산을 암호화하고 있었다. 그리고 한우 GLUT4 아미노산을 홀스타인, 사람, 생쥐와 비교한 결과 매우 높은 상동성을 나타내었다. 한우 각 조직에 있어서 GLUT4 mRNA의 발현을 확인한 결과 심장에서 가장 높은 발현을 나타내었으며 갈비, 등심, 대장에서는 낮은 발현을 보였다. 그리고 피하지방과 소장지방에서 GLUT4의 발현을 확인하기 위하여 RT-PCR을 수행한 결과 지방조직에서도 발현을 확인할 수 있었다. 한우 intramuscular preadipocyte 세포가 지방세포로 분화하는 과정에 있어서 GLUT4의 발현을 RT-PCR로 확인한 결과 분화에 따라 점차 줄어드는 경향을 나타내었다. 이와 같은 결과는 한우 지방조직에서의 GLUT4 기능은 사람과 생쥐에서의 기능과 다르다는 것을 나타낸다.

• Genomics & Informatics
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• 제15권4호
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• pp.162-169
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• 2017

Metal binding proteins or metallo-proteins are important for the stability of the protein and also serve as co-factors in various functions like controlling metabolism, regulating signal transport, and metal homeostasis. In structural genomics, prediction of metal binding proteins help in the selection of suitable growth medium for overexpression's studies and also help in obtaining the functional protein. Computational prediction using machine learning approach has been widely used in various fields of bioinformatics based on the fact all the information contains in amino acid sequence. In this study, random forest machine learning prediction systems were deployed with simplified amino acid for prediction of individual major metal ion binding sites like copper, calcium, cobalt, iron, magnesium, manganese, nickel, and zinc.

• BMB Reports
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• 제34권4호
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• pp.305-309
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• 2001

MatR in Rhizobium trifolii is a malonate-responsive transcription factor that regulates the expression of genes, matABC, enabling decarboxylation of malonyl-CoA into acetyl-CoA, synthesis of malonyl-CoA from malonate and CoA, and malonate transport. According to an analysis of the amino acid sequence homology, MatR belongs to the GntR family The proteins of this family have two-domain folds, the N-terminal helix-turn-helix DNA-binding domain and the C-terminal ligand-binding domain. In order to End the malonate binding site and amino acid residues that interact with RNA polymerase, a site-directed mutagenesis was performed. Analysis of the mutant MatR suggests that Arg-160 might be involved in malonate binding, whereas Arg-102 and Arg-174 are critical for the repression activity by interacting with RNA polymerase.

• Ocean and Polar Research
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• 제25권4호
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• pp.617-623
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• 2003

Organisms living in the Arctic and Antarctic regions are exposed to strong constraints, of which temperature is a driving factor. Evolution has led to special adaptations, some with important implications at the biochemical, physiological, and molecular levels. The northern and southern polar oceans have very different characteristics. Tectonic and oceanographic events have played a key role in delimiting the two polar ecosystems and influencing evolution. Antarctica has been isolated and cold longer than the Arctic; its ice sheet developed at least 10 million years earlier. As an intermediate system, the Arctic is a connection between the more extreme, simpler Antarctic system and the very complex temperate and tropical systems. By studying the molecular bases of cold adaptation in polar fish, and taking advantage of the information available on hemoglobin structure and function, we analysed the evolutionary history of the ${\alpha}\;and\;{\beta}globins$ of Antarctic and Arctic hemoglobin using the molecular clock hypothesis as a basis for reconstructing the phylogenetic relationships among species.