• Title/Summary/Keyword: Small protein

Search Result 1,755, Processing Time 0.034 seconds

Mitochondrial Genome Sequence of Echinostoma revolutum from Red-Crowned Crane (Grus japonensis)

  • Ran, Rongkun;Zhao, Qi;Abuzeid, Asmaa M.I.;Huang, Yue;Liu, Yunqiu;Sun, Yongxiang;He, Long;Li, Xiu;Liu, Jumei;Li, Guoqing
    • Parasites, Hosts and Diseases
    • /
    • v.58 no.1
    • /
    • pp.73-79
    • /
    • 2020
  • Echinostoma revolutum is a zoonotic food-borne intestinal trematode that can cause intestinal bleeding, enteritis, and diarrhea in human and birds. To identify a suspected E. revolutum trematode from a red-crowned crane (Grus japonensis) and to reveal the genetic characteristics of its mitochondrial (mt) genome, the internal transcribed spacer (ITS) and complete mt genome sequence of this trematode were amplified. The results identified the trematode as E. revolutum. Its entire mt genome sequence was 15,714 bp in length, including 12 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and one non-coding region (NCR), with 61.73% A+T base content and a significant AT preference. The length of the 22 tRNA genes ranged from 59 bp to 70 bp, and their secondary structure showed the typical cloverleaf and D-loop structure. The length of the large subunit of rRNA (rrnL) and the small subunit of rRNA (rrnS) gene was 1,011 bp and 742 bp, respectively. Phylogenetic trees showed that E. revolutum and E. miyagawai clustered together, belonging to Echinostomatidae with Hypoderaeum conoideum. This study may enrich the mitochondrial gene database of Echinostoma trematodes and provide valuable data for studying the molecular identification and phylogeny of some digenean trematodes.

Heterologous Expression of Yeast Prepro-$\alpha$-factor in Rat $GH_3$ Cells

  • Lee, Myung-Ae;Cheong, Kwang-Ho;Han, Sang-Yeol;Park, Sang-Dai
    • Animal cells and systems
    • /
    • v.4 no.2
    • /
    • pp.157-163
    • /
    • 2000
  • Yeast pheromone a-factor is a 13-amino acid peptide hormone that is synthesized as a part of a larger precursor, prepro-$\alpha$-factor, consisting of a signal peptide and a proregion of 64 amino acids. The carboxy-terminal half of the precursor contains four tandem copies of mature $\alpha$-factor. To investigate the molecular basis of intracellular sorting, proteolytic processing, and storage of the peptide hormone, yeast prepro-$\alpha$-factor precursors were heterologously expressed in rat pituitary $GH_3 cells. When cells harboring the precursor were metabolically labeled, a species of approximately 27 kD appeared inside the cells. Digestion with peptide: N-glycosidase F (PNG-F) shifted the molecular mass to a 19 kD, suggesting that the 27 kD protein was the glycosylated form as in yeast cells. The nascent polypeptide is efficiently targeted to the ER in the $GH_3 cells, where it undergoes cleavage of its signal peptide and core glycosylation to generate glycosylated pro-a-factor. To look at the post ER intracellular processing, the pulse-labelled cells were chased up to 2 hrs. The nascent propeptides disappeared from the cells at a half life of 30 min and only 10-25% of the newly synthesized, unprocessed precursors were stored intracellularly after the 2 h chase. However, about 20% of the pulse-labeled pro-$\alpha$-factor precursors were secreted into the medium in the pro-hormone form. With increasing chase time, the intracellular level of propeptide decreased, but the amount of secreted propeptide could not account for the disappearance of intracellular propeptide completely. This disappearance was insensitive to lysosomotropic agents, but was inhibited at $16^{circ}C or 20^{\circ}C$, suggesting that the turnover of the precursors was not occurring in the secretory pathway to trans Golgi network (TGN) or dependent on acidic compartments. From these results, it is concluded that a pan of these heterologous precursors may be processed at its paired dibasic sites by prohormone processing enzymes located in TGN/secretpry vesicles producing small peptides, and that the residual unprocessed precursors may be secreted into the medium rather than degraded intracellularly.

  • PDF

Role of Iridin Isolated from Iris koreana Nakai on Doxorubicin-induced Necrosis in HK-2 Cells, and Effect on Cancer Cells (노랑붓꽃에서 분리된 Iridin의 독소루비신 유도 HK-2 세포 괴사에 대한 역할 및 암세포에 대한 작용)

  • Nho, Jong Hyun;Lee, Ki Ho;Jung, Ho Kyung;Lee, Mu Jin;Jang, Ji Hun;Sim, Mi Ok;Jung, Ja Kyun;Jung, Da Eun;Cho, Hyun Woo
    • Korean Journal of Plant Resources
    • /
    • v.31 no.2
    • /
    • pp.95-101
    • /
    • 2018
  • Doxorubicin is a anti-cancer drugs that interferes with the growth and spread of cancer cells in human body. Doxorubicin is used to treat different types of cancers that affect the ovary, thyoid and lungs, but induced side effect such as nephrotoxicity and cardiotoxicity. Thus, we investigated that the effect of iridin on doxorubicin-induced necrosis in HK-2 cells, a human proximal tubule cell. To confirm effect of iridin on doxorubicin-induced necrosis, HK-2 cells are treated with $10{\mu}M$ doxorubicin and $80{\mu}M$ iridin. $80{\mu}M$ iridin reduced $10{\mu}M$ doxorubicin-induced necrosis, the mitochondrial over activation and caspase-3 activation. However, iridin reduces anti-cancer effect of doxorubicin such as PARP1 and caspase-3 activation, checkpoint proteins (CDK4 and CDK6) in NCI-H1129 cells (Human non-small cell lung cancer cell). In HCT-116 cells (Human colorectan cancer cell), iridin do not increased protein expression of CDK4 and CDK6 decreased by doxorubicin. Results indicate that treatment of iridin was diminished doxorubicin-induced necrosis in HK-2 cells. However, iridin was decreased anti-cancer effect of doxorubicin on NCI-H1229, but not HCT-116. Thus, further experiment are required to iridin treatment on various cancer cells and animal models because effect of iridin different cell type.

Metabolic Engineering for Resveratrol Derivative Biosynthesis in Escherichia coli

  • Jeong, Yu Jeong;Woo, Su Gyeong;An, Chul Han;Jeong, Hyung Jae;Hong, Young-Soo;Kim, Young-Min;Ryu, Young Bae;Rho, Mun-Chual;Lee, Woo Song;Kim, Cha Young
    • Molecules and Cells
    • /
    • v.38 no.4
    • /
    • pp.318-326
    • /
    • 2015
  • We previously reported that the SbROMT3syn recombinant protein catalyzes the production of the methylated resveratrol derivatives pinostilbene and pterostilbene by methylating substrate resveratrol in recombinant E. coli. To further study the production of stilbene compounds in E. coli by the expression of enzymes involved in stilbene biosynthesis, we isolated three stilbene synthase (STS) genes from rhubarb, peanut, and grape as well as two resveratrol O-methyltransferase (ROMT) genes from grape and sorghum. The ability of RpSTS to produce resveratrol in recombinant E. coli was compared with other AhSTS and VrSTS genes. Out of three STS, only AhSTS was able to produce resveratrol from p-coumaric acid. Thus, to improve the solubility of RpSTS, VrROMT, and SbROMT3 in E. coli, we synthesized the RpSTS, VrROMT and SbROMT3 genes following codon-optimization and expressed one or both genes together with the cinnamate/4-coumarate:coenzyme A ligase (CCL) gene from Streptomyces coelicolor. Our HPLC and LC-MS analyses showed that recombinant E. coli expressing both ScCCL and RpSTSsyn led to the production of resveratrol when p-coumaric acid was used as the precursor. In addition, incorporation of SbROMT3syn in recombinant E. coli cells produced resveratrol and its mono-methylated derivative, pinostilbene, as the major products from p-coumaric acid. However, very small amounts of pterostilbene were only detectable in the recombinant E. coli cells expressing the ScCCL, RpSTSsyn and SbROMT3syn genes. These results suggest that RpSTSsyn exhibits an enhanced enzyme activity to produce resveratrol and SbROMT3syn catalyzes the methylation of resveratrol to produce pinostilbene in E. coli cells.

Lipid composition of Korean safflower seeds (한국산 재래종 잇꽃 종실의 지질 성분)

  • Noh, Wan-Seob;Park, Jong-Sun
    • Applied Biological Chemistry
    • /
    • v.35 no.2
    • /
    • pp.110-114
    • /
    • 1992
  • The lipid compositions, physical and chemical characteristics of crude oil in Korean safflow, Carthamus tinctorius L., seeds were determined and proxmate composition of it were also analyzed. The proximate composition showed moisture 7.2%, carbohydrate 34.4%, crude protein 18.2%, crude fat 34.8% and ash 5.35. Specific gravity, refractive index, smoking point, melting point and titer of the crude oil were $0.922{\sim}0.927,\;1.468{\sim}1.469,\;210^{\circ}C,\;-15{\sim}-18^{\circ}C\;and\;15{\sim}18$, respectively. Iodine value, saponification value, acid value, esterification value, unsapoifiable matter and saturated fatty acid content of the crude oil were $140{\sim}152,\;186{\sim}192,\;2.6{\sim}3.5,\;179{\sim}186,\;0.8{\sim}1.3%\;and\;6.6{\sim}7.2%$, respectively. Fatty acid compositions were quantitatively determined by G.L.C. to give $70{\sim}78%$ linoleic acid, $10{\sim}23%$ oleic acid and $5{\sim}8%$ palmitic acid as main components, stearic, linolenic and myristic acid were presented in small quantities.

  • PDF

Generation and Characterization of a Neutralizing Human Monoclonal Antibody to Hepatitis B Virus PreS1 from a Phage-Displayed Human Synthetic Fab Library

  • Jo, Gyunghee;Jeong, Mun Sik;Wi, Jimin;Kim, Doo Hyun;Kim, Sangkyu;Kim, Dain;Yoon, Jun-Yeol;Chae, Heesu;Kim, Kyun-Hwan;Hong, Hyo Jeong
    • Journal of Microbiology and Biotechnology
    • /
    • v.28 no.8
    • /
    • pp.1376-1383
    • /
    • 2018
  • The hepatitis B virus (HBV) envelope contains small (S), middle (M), and large (L) proteins. PreS1 of the L protein contains a receptor-binding motif crucial for HBV infection. This motif is highly conserved among 10 HBV genotypes (A-J), making it a potential target for the prevention of HBV infection. In this study, we successfully generated a neutralizing human monoclonal antibody (mAb), 1A8 (IgG1), that recognizes the receptor-binding motif of preS1 using a phage-displayed human synthetic Fab library. Analysis of the antigen-binding activity of 1A8 for different genotypes indicated that it can specifically bind to the preS1 of major HBV genotypes (A-D). Based on Bio-Layer interferometry, the affinity ($K_D$) of 1A8 for the preS1 of genotype C was 3.55 nM. 1A8 immunoprecipitated the hepatitis B virions of genotypes C and D. In an in vitro neutralization assay using HepG2 cells overexpressing the cellular receptor sodium taurocholate cotransporting polypeptide, 1A8 effectively neutralized HBV infection with genotype D. Taken together, the results suggest that 1A8 may neutralize the four HBV genotypes. Considering that genotypes A-D are most prevalent, 1A8 may be a neutralizing human mAb with promising potential in the prevention and treatment of HBV infection.

Cloning and Characterization of Homeodomain-Zip Gene, Phc5 in Embryogenic Callus derived from Pimpinella brachycarpa Suspension Cultured Cells (참나물 현탁배양세포 유래 배발생캘러스에서 HD-Zip 유전자, Phc5의 클로닝과 특성)

  • 손수인;김준철
    • Korean Journal of Plant Tissue Culture
    • /
    • v.26 no.2
    • /
    • pp.121-126
    • /
    • 1999
  • Calli were induced from the petiole explants of Pimpinella brachycarpa on MS medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L BA after four weeks of culture. Compact clusters of small and dense cells among these calli were selected and suspension-cultured as the source of embryogenic calli. When transferred to MS medium with 0.1 mg/L NAA, the suspension-cultured cells grew to embryogenic callus. Somatic embryos derived from these embryogenic calli developed into plantlets. The cDNA library was constructed in the embryogenic callus and in order to screen the cDNA library, these cDNAs were plated at a density 1.5 $\times$ 10^5 plaques per 15 cm petridish. Among 19 clones showing preferential hybridization with petiole HD-Zip gene, five clones were obtained after second screening. Four clones among them, were highly homologous to P. brachycarpa shoot-tip Phz4 gene, but one clone, Phc5 was about 1.5 kb which has an extra 163 bp to 5' upstream of Phz4. The Phc5 was 1,531 bp containing poly A tails of 18 bases. ATG start codon for Phc5, was located at position 284 with an open reading frame of 906 by which encodes a polypeptide of 302 amino acids. The Phc5 protein revealed that the polypeptides between 135 and 195 contain a homeodomain as the `leucine zipper' motif.

  • PDF

Comparison of Functional Properties of Cheonggukjang by Using Red Ginseng (홍삼을 이용한 청국장의 기능적 특성 비교)

  • Park, Nan-Young;Seong, Jong-Hwon;Choi, Myung-Sook;Moon, Kwang-Deog;Kwon, Joong-Ho;Jeong, Yong-Jin
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.37 no.3
    • /
    • pp.261-268
    • /
    • 2008
  • To utilize Cheonggukjang as a raw material of diverse foods, the quality characteristics of Cheonggukjang and its processed food were compared. Cheonggukjang (CGJ), red ginseng Cheonggukjang (RCJ), and red ginseng Cheonggukjang hydrolysate (RCH) were powdered, and their quality and functional characteristics were examined. The result showed that in regard to general components, carbohydrate content of RCJ was higher than other samples while crude protein content was lower. Free amino acid content of RCH was 2,157.16 mg%, which was approximately 2 times higher than CGJ, and the content of essential amino acid was 812.18 mg%, which was the highest. The result of SDS-electrophoresis pattern showed that CGJ and RCJ showed a molecular weight smaller than 33 kDa, and RCH showed a smaller than 17 kDa low molecular weight, confirming the hydrolysis to small molecular weight. Among the samples, free radical scavenging activity, superoxide radical scavenging activity, and ACE inhibitory activity did not show a significant difference; nonetheless, RCH showed the highest activity while CGJ showed the highest fibrinolytic activity of 111.38 unit. In addition, in sensory evaluation, the peculiar bitter taste of red ginseng could be detected while the overall acceptability was improved. Based on the above results, in comparison with CGJ, as for RCJ and RCH, their function was strengthened and unpleasant odor was reduced, and thus it is anticipated that they could be used as a raw material of diverse foods.

Analysis on the Components of the Elaeagnus multiflora Thunb. Leaves (뜰보리수 잎의 유용성분 분석)

  • Yoon, Kyung-Young;Hong, Ju-Yeon;Shin, Seung-Ryeul
    • Food Science and Preservation
    • /
    • v.14 no.6
    • /
    • pp.639-644
    • /
    • 2007
  • This study analyzed components of the leaves of Elaeagnus multiflora as part of studies on the nutritional and functional materials of fruits and leaves of this plant. The moisture content of the leaves was 71.6% and the carbohydrate, crude protein, lipid and ash contents were 24.1, 1.4, 0.4 and 2.5%, respectively. Concentrations of reducing sugars, soluble proteins and polyphenols were 460.0, 503.3 and 805.6 mg/100 g, respectively. Fructose was the dominant free sugar, and arabinose, maltose, glucose, and a small amount of trehalose were also detected. Malic acid was the main organic acid in E. multiflora leaves, and acetic acid, citric acid, lactic acid, and succinic acid were also present. E. multiflora leaves were high in K, Ca and Mg. Of hydrolyzed amino acids, alanine was present at the highest concentration (112.0 mg/100 g), with threonine, leucine, valine and phenylalanine being the next most common. Glutamic acid and ornithine were the dominant free amino acid and amino acid derivative, respectively.

Pharmacokinetics of Recombinant Human Epidermal Growth Factor (DWP401) after Repeated Subcutaneous Administration to Rats (흰쥐에 재조합 인간 상피세포 성장인자(DWP401)를 연용피하투여했을 때 약물체내동태)

  • Nam, Gwon-Ho;Jo, Jae-Yeol;Jeong, Ju-Yeong;Jang, U-Ik;Gang, Jin-Seok;Yu, Eun-Suk;Park, Seung-Guk;Yu, Yeong-Hyo;Park, Myeong-Hwan;Sim, Chang-Gu
    • YAKHAK HOEJI
    • /
    • v.40 no.5
    • /
    • pp.491-500
    • /
    • 1996
  • The organ distribution and pharmacokinetics of DWP401, a recombinant human epidermal growth factor (rhEGF), were compared after single and repeated subcutaneous administration ( 50${\mu}$/kg, 10${\mu}g$Ci/kg of $^{125}I$-DWP401, twice a day for 7 consecutive days) to rats. The pharmacokinetic parameters such as AUC and terminal half-life were similar between two different administration. During repeated administration, the plasma concentration of DWP401 seemed to be constant when the plasma was collected at 15 min after each dosing. The TCA-precipitated radioactivities in thyroid, liver, kidney, and stomach were higher than those of other organs studied after both single and repeated administration. The TCA-precipitated radioactivities after repeated administration in several organs, such as thyroid, stomach, prostate, adrenal, eye ball, and testis were higher than those after single administration. But, according to the observations using gel filtration chromatography and antibody binding assay, the radioactivities in thyroid and stomach were not primarily due to the intact DWP401 or its metabolites but due to the $^{125}I$-thyroxine binding protein. In conclusion, it can be suggested that DWP401 is metabolized to each amino acid or small polypeptides, and there was no significant changes in pharmacokinetics or any indications for accumulation of DWP401 in rat plasma and organs after repeated treatment.

  • PDF