• Title/Summary/Keyword: Small protein

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Interaction between a Blood Vessel-Inducing Protein Angiogenin and Its Binding Protein Actin

  • Chang, Soo-Ik;Paik, Seung-Bum;So, Seung-Ho;Ahn, Byung-Cheol
    • BMB Reports
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    • v.29 no.4
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    • pp.353-358
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    • 1996
  • Bovine angiogenin (bAng) is a potent blood vessel inducing protein purified from cow In ilk. fluorescence spectroscopy has been used to study the interaction of bAng with actin in 50 mM Tris-HCl pH 7.5, and 1 mM $CaCl_2$ at $25^{\circ}C$. Actin contains four tryptophans but bAng contains no tryptophans. A 50% decrease in intrinsic fluorescence accompanied formation of the bAng/actin complex. By contrast, the interaction of RNase A, a homologous protein to bAng, with actin results in about 10% quenching of the fluorescence. Fluorescence titration experiments were performed by adding increasing concentrations of bAng (0~1.0 ${\mu}M$) to a constant concentration of actin (0.1 ${\mu}M$), and the dissociation constant $K_d$ for the bAng/actin complex and the stoichiometry n were measured as $20{\pm}1$ nM and $1.0{\pm}0.1$ respectively. These results suggest that the interaction between bAng with actin is specific and that quenching of actin fluorescence has occurred in the bAng/actin complex. The bAng binding sites of actin are discussed in the results of this study, and we propose that Trp-80 in the small domain of bovine actin is responsible for the bAng/actin binding.

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Polyhydroxyalkanoate Chip for the Specific Immobilization of Recombinant Proteins and Its Applications in Immunodiagnostics

  • Park, Tae-Jung;Park, Jong-Pil;Lee, Seok-Jae;Hong, Hyo-Jeong;Lee, Sang-Yup
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.173-177
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    • 2006
  • In this study, a novel strategy was developed for the highly selective immobilization of proteins, using the polyhydroxyalkanoate (PHA) depolymerase substrate binding domain (SBD) as an active binding domain. In order to determine the appropriacy of this method for immunodiagnostic assays, the single-chain antibody (ScFv) against the hepatitis B virus (HBV) preS2 surface protein and the severe acute respiratory syndrome coronavirus (SARS-CoV) envelope protein (SCVe) were fused to the SBD, then directly immobilized on PH A-coated slides via microspotting. The fluorescence-labeled HBV antigen and the antibody against SCVe were then utilized to examine specific interactions on the PHA-coated surfaces. Fluorescence signals were detected only at the spotted positions, thereby indicating a high degree of affinity and selectivity for their corresponding antigens/antibodies. Furthermore, we detected small amounts of ScFv-SBD (2.7 ng/mL) and SCVe-SBD fusion proteins (0.6ng/mL). Therefore, this microarray platform technology, using PHA and SBD, appears generally appropriate for immunodiagnosis, with no special requirements with regard to synthetic or chemical modification of the biomolecules or the solid surface.

CYP1B1 Activates Wnt/β-Catenin Signaling through Suppression of Herc5-Mediated ISGylation for Protein Degradation on β-Catenin in HeLa Cells

  • Park, Young-Shin;Kwon, Yeo-Jung;Chun, Young-Jin
    • Toxicological Research
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    • v.33 no.3
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    • pp.211-218
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    • 2017
  • Cytochrome P450 1B1 (CYP1B1) acts as a hydroxylase for estrogen and activates potential carcinogens. Moreover, its expression in tumor tissues is much higher than that in normal tissues. Despite this association between CYP1B1 and cancer, the detailed molecular mechanism of CYP1B1 on cancer progression in HeLa cells remains unknown. Previous reports indicated that the mRNA expression level of Herc5, an E3 ligase for ISGylation, is promoted by CYP1B1 suppression using specific small interfering RNA, and that ISGylation may be involved in ubiquitination related to ${\beta}-catenin$ degradation. With this background, we investigated the relationships among CYP1B1, Herc5, and ${\beta}-catenin$. RT-PCR and western blot analyses showed that CYP1B1 overexpression induced and CYP1B1 inhibition reduced, respectively, the expression of $Wnt/{\beta}-catenin$ signaling target genes including ${\beta}-catenin$ and cyclin D1. Moreover, HeLa cells were treated with the CYP1B1 inducer $7,12-dimethylbenz[{\alpha}]anthracene$ (DMBA) or the CYP1B1 specific inhibitor, tetramethoxystilbene (TMS) and consequently DMBA increased and TMS decreased ${\beta}-catenin$ and cyclin D1 expression, respectively. To determine the correlation between CYP1B1 expression and ISGylation, the expression of ISG15, a ubiquitin-like protein, was detected following CYP1B1 regulation, which revealed that CYP1B1 may inhibit ISGylation through suppression of ISG15 expression. In addition, the mRNA and protein expression levels of Herc5 were strongly suppressed by CYP1B1. Finally, an immunoprecipitation assay revealed a direct physical interaction between Herc5 and ${\beta}-catenin$ in HeLa cells. In conclusion, these data suggest that CYP1B1 may activate $Wnt/{\beta}-catenin$ signaling through stabilization of ${\beta}-catenin$ protein from Herc5-mediated ISGylation for proteosomal degradation.

An Indigenous Case of Intestinal Capillariasis with Protein-Losing Enteropathy in Korea

  • Jung, Woon Tae;Kim, Hyun Jin;Min, Hyun Ju;Ha, Chang Yoon;Kim, Hong Jun;Ko, Gyung Hyuck;Na, Byoung-Kuk;Sohn, Woon-Mok
    • Parasites, Hosts and Diseases
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    • v.50 no.4
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    • pp.333-337
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    • 2012
  • We encountered an indigenous case of intestinal capillariasis with protein-losing enteropathy in the Republic of Korea. A 37-year-old man, residing in Sacheon-si, Gyeongsangnam-do, admitted to the Gyeongsang National University Hospital (GNUH) due to long-lasting diarrhea, abdominal pain, anasarca, and weight loss. He recalled that he frequently ate raw fish, especially the common blackish goby (Acanthogobius flavimanus) and has never been abroad. Under the suspicion of protein-losing enteropathy, he received various kinds of medical examinations, and was diagnosed as intestinal capillariasis based on characteristic sectional findings of nematode worms in the biopsied small intestine. Adults, juvenile worms, and eggs were also detected in the diarrheic stools collected before and after medication. The clinical symptoms became much better after treatment with albendazole 400 mg daily for 3 days, and all findings were in normal range in laboratory examinations performed after 1 month. The present study is the 6th Korean case of intestinal capillariasis and the 3rd indigenous one in the Republic of Korea.

Expression of the Heat Shock Protein Genes in Response to Thermal Stress in the Silkworm Bombyx mori

  • Velu, Dhanikachalam;Ponnuvel, Kangayam. M.;Qadri, Syed. M. Hussaini
    • International Journal of Industrial Entomology and Biomaterials
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    • v.16 no.1
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    • pp.21-27
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    • 2008
  • The expression of heat shock protein genes (Hsp 70, Hsp 40, Hsp 20.8 and Hsp 20.4) against thermal stress in silkworm Bombyx mori was performed through semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Upon exposure of silkworm to two temperature regimes ($38^{\circ}C$ and $42^{\circ}C$), significant change in the expression of Hsp gene was observed as compared to the control. Hsp 70 and Hsp 40 showed increased expression than the small heat shock protein genes Hsp 20.8 and Hsp 20.4. The Hsp 70 showed increased expression during the recovery period as compared to 1 hr thermal treatments ($38^{\circ}C$/1 hr and $42^{\circ}C$/1 hr). Whereas, Hsp 40, Hsp 20.8 and Hsp 20.4 genes showed higher expression level at initial stages that later gradually decrease during recovery period. Tissue specific expression of Hsp 70 showed variation in the level of expression amongst the tissues. The mid gut and fat body tissues showed higher expression than the cuticle and silk gland tissue. The Hsp 70, Hsp 40 gene expression was analyzed in thermotolerant (Nistari) and thermo susceptible silk worm strain (NB4D2) and results showed significant variation in their expression level. The Nistari showed higher expression of Hsp 70 and Hsp 40 genes than the NB4D2. These findings provide a better understanding of cellular protection mechanisms against environmental stress such as heat shock, as these Hsps are involved in an organism thermotolerance.

The effect of some detergents on the changes of bacterial membrane (계면 활성제 처리에 의한 세균 세포막의 변화에 관한 연구)

  • 이종삼;이호용;조기승;조선희;장성열;최영길
    • Korean Journal of Microbiology
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    • v.21 no.3
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    • pp.115-126
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    • 1983
  • The results that the effect of 6 detergents on the structural changes and biochemical composition of bacterial membrane of Escherichia coli and Bacillus cereus are as follows ; 1. Population growth of the bacteria was increased in case of the treatment with palmitoyl carnitine and sodium deoxy cholate but was increased in case of the treatment with palmitoyl carnitine and sodium deoxy cholate but was decreased by sodium dodecyl sulfate and palmitoyl choline, in E.coli and was decreased by palmitoyl carnitine and palmitoyl choline at the low concentration, in B. cereus. 2. The electron micrograph showed that cell wall lysis or cell collapse were observed in the treatment of sodium dodecyl sulfate and palmitoyl choline, and also cell wall was condensed by triton X-100 and sodium deoxy cholate, in E.coli. And in B. cereus, endospore formation of the bacteria was stimulated by palmitoyl choline, and cell lysis or structural changes of the membrane were observed in the treatment of sodium dodecyl sulfate, sodium cholate, and triton X-100, respectively. 3. As to the effect of detergent on the biochemical composition of biomembrane, the content of carnitine, in E.coli, and B.cereus, the content of structural protein and phospholipid were decreased by treatment of sodium dodecyl sulfate and structural protein was denatured by palmitoyl choline. 4. The profile of membrane protein revealed that the bacterial membrane were composed of various proteins. By dint of this result, some of membrane proteins were solubilized or changed to small molecules by the treatment of sodium dodecyl sulfate and palmitoyl choline, in E.coli and membrane protein of the biomembrane by treatment of sodium dodecyl sulfate, sodium deoxy cholate, palmitoyl choline, and palmitoyl carnitine were confirmed to be different profile as compared with those of the control, in B. cereus. Therefore, it is suggested that sodium dfodecyl sulfate and palmitoyl choline soulbilized biomembranes or inhibited membrane transport and that palmitoyl carnitine and sodium deoxy cholate were used as an energy source or stimulating the membrane transport, in E.coli. And, it is suggested that all of detergents were inhibited biomembrane synthesis, expet saponin, in B.cereus.

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Chemical Composition of Seed in Medicinal Soybean Collected in Korea

  • Seong, Rak-Chun;Kim, Jeong-Gyu;Hwang, Young-Hyun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.3
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    • pp.141-146
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    • 1998
  • Production of medicinal soybean [Glycine max (L.) Merrill], characterized with black seed, white stripe at hilum border, yellow cotyledon and small seed, is increasing with increasing consumption. The objective of this study was to investigate the chemical composition of medicinal soybean seed and to provide basic information, for the characterization of these soybeans among genetic resources. Forty-four lines of medicinal soybeans collected from Korea and two control cultivars, 'Hwangkeumkong' (Yellow seed coat) and 'Geomjeongkong l' (Black seed coat) were planted at the Research Farm of the College of Natural Resources, Korea University, located at Namyangju City on May 25, 1996. Seeds of these lines were harvested at full maturity and analyzed for protein, oil, sugar, starch and mineral contents. Mean protein and oil content of the medicinal line seeds were 42.6 and 16.1%, respectively, and those of the control cultivars were in the middle range for protein and oil content. However, sugar and starch content of the medicinal line seeds appeared to be in the lower range of the distributions compared to the control cultivars and were 10.0 and 1.68%, respectively. Mean P, K, Ca, and Mg contents of the seeds of medicinal soybean lines were 15.9, 21.5, 3.11, and 2.81 mg/g, respectively, indicating that these lines had higher P, K, and Mg and lower Ca contents when compared to the control cultivars. Mean Na and Fe contents were 671 and 224 mg/kg, respectively, showing lower Na and similar Fe contents. The observed results provided that chemical compositions of medicinal soybean were, on average, different from those of the general soybean cultivars.

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Nutrition survey of the Aged on Jeju Island (제주지역 고령자 영양실태 조사연구)

  • 고양숙
    • Journal of the Korean Home Economics Association
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    • v.19 no.4
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    • pp.41-53
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    • 1981
  • The purpose of this survey was to investigate the nutrient and food intake and food habits of the aged Jeju island. The survey was conducted to 102 persons, 50 persons aged 65 to 74 years and 52 persons aged 75 years over from July 5 to 25, 1981. The result obtained was summarized as follows; 1. Average nutrient intake per day Calorie intake was 1408 ㎉(which was 64% of RDA) for male and was 1343 ㎉ (84%) for female aged 65 to 74 years. For male aged 75 years over calorie intake was 1316㎉ (60%) and for female was 1292㎉ (81%). According to the intake of calorie, the ratio of carbohydrate, protein, fat, were average 74.8%, 15.1%, 10.1%. Protein intake was insufficient quantiatively, but the proportion of animal protein to total protein intake were 26.1 to 33.5%. Other nutrient intake, such as Ca, vitamin A and C were lower than the recommended dietary allowance. 2. Food intake In per capita food consumption were 69.1g to 744.7g and average 320.1g of cereal (125.1g of rice and 160.5g of barley) and 63.9g of fishes. Total food consumption and vegetables were diminished by older age. 3. Correlation assessment The correlation coefficient between residental stus lived with family, high activity, good health, and nutrient intake were significant (P<0.05). Poor teeth had influence on less nutrient intake, but was not significant. 4. Food preferences Food liked over 70% of subjects were persimmon, fishes, leef, tomato, and boiled rice with barley, etc. owing to tender food, economical reasons, seasonal influence, and food habits. Food disliked over 20% of subjects were instant noodle, small sardin, boiled rice, and ice-crea, etc. The majority of subjects had never had the processing food, such as ham, sausage, butter and margarine, etc.

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IQGAP1, a signaling scaffold protein, as a molecular target of a small molecule inhibitor to interfere with T cell receptor-mediated integrin activation

  • Li, Lin-Ying;Nguyen, Thi Minh Nguyet;Woo, Eui Jeon;Park, Jongtae;Hwang, Inkyu
    • Korean Journal of Agricultural Science
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    • v.47 no.2
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    • pp.361-373
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    • 2020
  • Integrins such as lymphocyte function-associated antigen -1 (LFA-1) have an essential role in T cell immunity. Integrin activation, namely, the transition from the inactive conformation to the active one, takes place when an intracellular signal is generated by specific receptors such as T cell receptors (TCRs) and chemokine receptors in T cells. In an effort to explore the molecular mechanisms underlying the TCR-mediated LFA-1 activation, we had previously established a high-throughput cell-based assay and screened a chemical library deposited in the National Institute of Health in the United States. As a result, several hits had been isolated including HIKS-1 (Benzo[b]thiophene-3-carboxylic acid, 2-[3-[(2-carboxyphenyl) thio]-2,5-dioxo-1-pyrrolinyl]-4,5,6,7-tetrahydro-,3-ethyl ester). In an attempt to reveal the mode of action of HIKS-1, in this study, we did drug affinity responsive target stability (DARTS) assay finding that HIKS-1 interacted with the IQ motif containing GTPase activating protein 1 (IQGAP1), a 189 kDa multidomain scaffold protein critically involved in various signaling mechanisms. Furthermore, the cellular thermal shift assay (CETSA) provided compelling evidence that HIKS-1 also interacted with IQGAP1 in vivo. Taken together, it can be concluded that HIKS-1 interferes with the TCR-mediated LFA-1 activation by interacting with IQGAP1 and thereby disrupting the signaling pathway for LFA-1 activation.

Relationship of the Signal Transduction-mediated Proteins and Enzymes to Contractility and Plasticity in Skeletal Muscles (골격근의 수축과 가소성에 대한 신호전달-매개 단백질 및 관련 효소의 상관성)

  • Kim, Jung-Hwan
    • The Journal of Korean Physical Therapy
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    • v.19 no.4
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    • pp.1-14
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    • 2007
  • Background: It is generally accepted that skeletal muscle contraction is triggered by nerve impulse and intracellular $Ca^{2+}\;([Ca^{2+}]_i)$ released from intracellular $Ca^{2+}$ stores such as sarcoplasmic reticulum (SR). Specifically, this process, called excitation-contraction (E-C) coupling, takes place at intracellular junctions between the plasma membrane, the transverse (T) tubule L-type $Ca^{2+}$ channel (dihydropyridine-sensitive L-rype $Ca^{2+}$ channel, DHPR, also called tetrads), and the SR $Ca^{2+}$ release channel (ryanodine-sensitive $Ca^{2+}$ release channel, RyR, also called feet) of internal $Ca^{2+}$ stores in skeletal muscle cells. Furthermore, it has been reported that the $Ca^{2+-}$ dependent and -independent contraction determine the expression of skeletal muscle genes, thus providing a mechanism for tightly coupling the extent of muscle contraction to regulation of muscle plasticity-related excitation-transcription (E-T) coupling. Purpose: Expression and activity of plasticity-associated enzymes in gastrocnemius muscle strips have not been well studied, however. Methods: Therefore, in this study the expression and phosphorylation of E-C and E-T coupling-related mediators such as protein kinases, ROS(reactive oxygen species)- and apoptosis-related substances, and others in gastrocnemius muscles from rats was examined. Results: I found that expression and activity of MAPKs (mitogen-activated protein kinases, ERK1/2, p38MAPK, and SAPK/JNK), apoptotic proteins (cleaved caspase-3, cytochrome c, Ref-1, Bad), small GTP-binding proteins (RhoA and Cdc42), actin-binding protein (cofilin), PKC (protein kinase C) and $Ca^{2+}$ channel (transient receptor potential channel 6, TRPC6) was observed in rat gastrocnemius muscle strips. Conclusion: These results suggest that MAPKs, ROS- and apoptosis-related enzymes, cytoskeleton-regulated proteins, and $Ca^{2+}$ channel may in part functionally import in E-C and E-T coupling from rat skeletal muscles.

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