• Title/Summary/Keyword: Stable protein 1

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Gamma-Irradiation Enhances RECK Protein Levels in Panc-1 Pancreatic Cancer Cells

  • Kim, Na Young;Lee, Jung Eun;Chang, Hyeu Jin;Lim, Chae Seung;Nam, Deok Hwa;Min, Bon Hong;Park, Gil Hong;Oh, Jun Seo
    • Molecules and Cells
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    • v.25 no.1
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    • pp.105-111
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    • 2008
  • Radiotherapy is an important treatment for many malignant tumors, but there are recent reports that radiation may increase the malignancy of cancer cells by stimulating expression of type IV collagenases. In this study, we examined changes in matrix metalloproteinase (MMP) inhibitors, such as the tissue inhibitors of metalloproteinase (TIMP)-1, TIMP-2 and RECK, in response to irradiation in Panc-1 pancreatic cancer cells. Irradiation increased RECK protein levels but not mRNA levels, whereas no significant changes were found in TIMP-1 and TIMP-2. The enhanced RECK protein levels were associated with an increase in MMP inhibitory activity. However, irradiation slightly but reproducibly increased the invasiveness of the Panc-1 cells. Like irradiation, treatment of Panc-1 cells with transforming growth factor $(TGF)-{\beta}1$ led to a 2-fold increase in RECK protein levels. Transient transfection with Smad3 also increased RECK protein levels, but transfection with Smad7 markedly reduced them. Stable expression of Smad7 and treatment with SB431542, an inhibitor of $TGF-{\beta}$ receptor I kinase, abolished $TGF-{\beta}1$- and radiation-mediated effects on RECK. Furthermore, irradiation increased levels of phosphorylated Smad3. We conclude that radiation post-transciptionally enhances RECK protein levels in Panc-1 cells, at least in part, via $TGF-{\beta}$ signaling, and that irradiation increases Panc-1 invasiveness via a mechanism that may not be linked to MMP-2 activity.

A Study on the Functional Properties of Camellia(Camellia japonica L.) Seed Protein Isolate (분리 동백단백의 기능적 특성)

  • 강성구
    • Korean Journal of Plant Resources
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    • v.11 no.3
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    • pp.272-278
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    • 1998
  • This study was carried out to investigate the functional properties such as nitrogen solubility, emulsifying property , foaming capapcity , water and oil absorption of Camellia (Camellia japonica .) seed protein isolate in condition of distilled water and 0.5M NaCl solution at pH 2.0∼10.0. Nitrogen solubility of Camellia protein isolate in distilled water showed the minimum value at pH 4.0 and increased at pH lower or higher than the isoelectric point(pH 4.0). It was 90.0 %at pH 10.0 Nitrogen solubility of 0.5M NaCl solution showed a similar pattern with that of distrille dwater but was higher than that of distilled water except pH 2.0 and pH 10.0. Emulsifying activity of Camellia seed protein islate showed the minimum value at pH 4.0, but was higher at ether value of pH. Emulsifying stability of protein isolate was stable by heat treatment for 30min, at 80℃ and increased in 0.5M NaCl solution more than that of distille dwater. Foaming capacity of Camellia seed protein isolate in distill3ed water showed the minimum value near the isoelectric point, While it changed little at other values of pH. Foaming stability slowly decreased as, but didn't make a significant difference as time was delayed . Oil absorption was 1.4ml per a sample of 1g and water absorption was 0.9ml per a sample of 1g. The former was higher than the latter . The content of total amino acid of Camellia protein isolate was 43.67% and the major total amino acid of Camellia protein isolate was 43.67% and the major total amino acid was in the order of glutamic acid , arginine, aspartic acid, and leucine.

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Induction of Boiling Stable Proteins by Cold and ABA Treatment in Radish Cotyledon and Hypocotyl (무 자엽과 하배축에서 저온과 ABA처리로 유도된 중탕에 강한 단백질 분석)

  • Cho, Bong-Heuy
    • Analytical Science and Technology
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    • v.13 no.3
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    • pp.346-350
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    • 2000
  • Boiling stable proteins of 53 kDa and 29 kDa existed natively in the cotyledons of Bak Kyoung, fall radish (Raphanus raphanistrodes L.) Boiling stable proteins of 36 kDa and 16.5 kDa were newly induced by cold stress and the proteins of 53 kDa and 29 kDa increased during the cold stress. The proteins of 53 kDa were denatured within 2 hrs after removing cotyledons from plants. Boiling stable proteins of 53 kDa existed natively in the hypocotyls as much as in the cotyledons whereas 24 kDa and 18 kDa proteins were increased by stress. Boiling stable proteins of 53 kDa were induced and those of the 25 kDa and 23 kDa were increased by cold treatment and ABA treatment in the cotyledons of Jangchundaehyung F1 spring white (Raphanus raphanistrodes L.). These results showed the differences of induced boiling stable proteins between fall radishes and spring radishes. Cycloheximide inhibited the induction of 25 kDa and 23 kDa proteins during stress. 22 kDa native protein disappeared during ABA treatment and reappeared by cycloheximide treatments. It may be explained that cycloheximide was responsible for the destruction process of proteins in the living organisms. The profile of boiling stable proteins in hypocotyls of spring radishes during stress was same as that of fall redishes.

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A bioinformatics approach to characterize a hypothetical protein Q6S8D9_SARS of SARS-CoV

  • Md Foyzur Rahman;Rubait Hasan;Mohammad Shahangir Biswas;Jamiatul Husna Shathi;Md Faruk Hossain;Aoulia Yeasmin;Mohammad Zakerin Abedin;Md Tofazzal Hossain
    • Genomics & Informatics
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    • v.21 no.1
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    • pp.3.1-3.10
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    • 2023
  • Characterization as well as prediction of the secondary and tertiary structure of hypothetical proteins from their amino acid sequences uploaded in databases by in silico approach are the critical issues in computational biology. Severe acute respiratory syndrome-associated coronavirus (SARS-CoV), which is responsible for pneumonia alike diseases, possesses a wide range of proteins of which many are still uncharacterized. The current study was conducted to reveal the physicochemical characteristics and structures of an uncharacterized protein Q6S8D9_SARS of SARS-CoV. Following the common flowchart of characterizing a hypothetical protein, several sophisticated computerized tools e.g., ExPASy Protparam, CD Search, SOPMA, PSIPRED, HHpred, etc. were employed to discover the functions and structures of Q6S8D9_SARS. After delineating the secondary and tertiary structures of the protein, some quality evaluating tools e.g., PROCHECK, ProSA-web etc. were performed to assess the structures and later the active site was identified also by CASTp v.3.0. The protein contains more negatively charged residues than positively charged residues and a high aliphatic index value which make the protein more stable. The 2D and 3D structures modeled by several bioinformatics tools ensured that the proteins had domain in it which indicated it was functional protein having the ability to trouble host antiviral inflammatory cytokine and interferon production pathways. Moreover, active site was found in the protein where ligand could bind. The study was aimed to unveil the features and structures of an uncharacterized protein of SARS-CoV which can be a therapeutic target for development of vaccines against the virus. Further research are needed to accomplish the task.

PKC-Independent Stimulation of Cardiac $Na^+/Ca^{2+}$ Exchanger by Staurosporine

  • Kang, Tong-Mook
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.5
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    • pp.259-265
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    • 2008
  • $[Ca^{2+}]_i$ transients by reverse mode of cardiac $Na^+/Ca^{2+}$ exchanger (NCX1) were recorded in fura-2 loaded BHK cells with stable expression of NCX1. Repeated stimulation of reverse NCX1 produced a long-lasting decrease of $Ca^{2+}$ transients ('rundown'). Rundown of NCX1 was independent of membrane $PIP_2$ depletion. Although the activation of protein kinase C (PKC) was observed during the $Ca^{2+}$ transients, neither a selective PKC inhibitor (calphostin C) nor a PKC activator (PMA) changed the degrees of rundown. By comparison, a non-specific PKC inhibitor, staurosporine (STS), reversed rundown in a dose-dependent and reversible manner. The action of STS was unaffected by pretreatment of the cells with calphostin C, PMA, or forskolin. Taken together, the results suggest that the stimulation of reverse NCX1 by STS is independent of PKC and/or PKA inhibition.

Nitrogen Compounds of Korea Ginseng and their Physiological Significance

  • Park, Hoon;Cho, Byung-Goo;Lee, Mee-Kyoung
    • Proceedings of the Ginseng society Conference
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    • 1990.06a
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    • pp.175-189
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    • 1990
  • Nitrogen compounds of Panax ginseng and their biological activities in plant and animal were reviewed. Major nitrogen compounds found in P. ginseng are free amino acids. Water solilble proteins, indouble proteins and peptides. Minor nitrogen compounds are dencichine. Glycolyroteins, amines, alkaloides, methoxy or alkyl pyrazine derivatives, free nucleosides and nucleic acid bases. 4-methyl-i-thiazoltethanol and pyroglutamic acid the contents of total nitrogen and protein in root Increased until 13 years old which was the highest age tinder investigation. Soluble protein content increased with the root weight and was higher in xylem pith than cortex-epidermis indicating the close relation with root growth. Arginine, which covered 58% of total free amino acids, may serve as storage nitrogen. Arginine seems to be changed into proline in rhizome. threonine in stem and again threonine and arginine in leaf. The greater the root weight the higher the polyamine stimulated Polyamine stimlllated the growth of root callus. Physiological roles of other minor nitrogen compounds are unknown although content is relatively high ((1.if) 6.w). Biochemical and pharmacological activities of some nitrogen compounds for animal were more investigated than physiological role there plant itself. Radiation and U.V protective function (heat stable protein). insulin-like activity in lipogenesis and livolysis (adenosine and pyroglutamic acid), depression of blood sugar content (glycopevtide). htmostatic and nellrotoxic activity (dencichine) and, sedative and hypnotic activity (4-methyl-i-thiazoleethanol) are reported. Heat stable protein increased with root age. The traditional quality criteria appear to be well in accordance with biological activities of nitrogen compounds. Chemical studies of nitrogen compounds seem relatively rare, probably due to difficulty of isolation, subsequently the investigations of biological activities are little.

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Complete Nucleotide Sequence of Tobacco Mosaic Virus Isolated from Wasabi(Eutrema wasabi Maxim.) (고추냉이에서 분리한 담배 모자이크 바이러스(TMV-W)의 전체 유전자 염기서열 분석)

  • 이귀재
    • Korean Journal of Plant Resources
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    • v.16 no.1
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    • pp.82-88
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    • 2003
  • Genomic RNA sequence of a tobamovirus infecting Eutrema wasabi plant(TMV-W) was determined. The RNA is composed 6,298 nucleotide and contains four OREs encoding the protein of 180KD(OREI), 130KD(ORE2),30KD(ORF3) and 18KD(coat protein, ORF4). ORE4, ORF 3, ORF 2 and ORF 1 are overlaped by 130, 20 and 40 nucleotides, and the overapping region can be folded into a stable hairpin styucture. This includes the 3'non-coding region of 238 nucleotides, coat protein gene(537 nucleotides,179 amino acid), 30KD movement protein gene(825 nucleotides, 275 amino acid), 13(IKD protein gene(1,896 nucleotides, 632 amino acid) and 180KD protein gene(2,958 nucleotides, 986 amino acid). The genomic RNA sequence was compared with homologous regions of eleven other tobamoviruses. TMV-WTE was similar to TMV-WSF(98.6%) in nucleotide sequence.

Intracellular Localization and Sustained Prodrug Cell Killing Activity of TAT-HSVTK Fusion Protein in Hepatocelullar Carcinoma Cells

  • Cao, Limin;Si, Jin;Wang, Weiyu;Zhao, Xiaorong;Yuan, Xiaomei;Zhu, Huifen;Wu, Xiaolong;Zhu, Jianzhong;Shen, Guanxin
    • Molecules and Cells
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    • v.21 no.1
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    • pp.104-111
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    • 2006
  • Gene therapy with nonviral vectors using the suicide gene/prodrug activating system of herpes simplex virus type-1 thymidine kinase (HSV1-TK)/ganciclovir (GCV) is inefficient in killing malignant tumor cells due to two major factors: (a) an unsatisfactory bystander effect; (b) short-lived expression of the protein. To study the capacity of the protein transduction domain (PTD) of HIV-1 TAT protein to enhance HSV1-TK/GCV cancer gene therapy, we constructed three fusion proteins TAT-TK, TK-TAT and TK. TAT-TK retained as much enzyme activity as TK, whereas that of TK-TAT was much lower. TAT-TK can enter HepG2 cells and much of it is translocated to the nucleus. The transduced HepG2 cells are killed by exogenously added GCV and have bystander effects on untransduced HepG2 cells. Most importantly, the introduced recombinant protein is stable and remains functional for several days at least, probably because nuclear localization protects it from the cytoplasmic degradation machinery and provides access to the nuclear transcription machinery. Our results indicate that TAT fusion proteins traffic intercellularly and have enhanced stability and prodrug cell killing activity. We conclude that TAT has potential for enhancing enzyme prodrug treatment of liver cancers.

Purification and Structural Characterization of P93 Complex from Hyperthermophilic Archaeon Thermococcus profundus (초고온 archaeon인 Thermococcus profundus에서 P93 복합체의 분리 및 구조적 특성)

  • Lee, Mi-Hong;Kim, Suk-Kyoung;Yun, Young-Gun;Park, Seong-Cheol;Bahk, Jeong-Dong;Cheong, Gang-Won
    • Applied Microscopy
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    • v.30 no.2
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    • pp.185-191
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    • 2000
  • An unusually large protein complex was found in the cytosol of the hyperthmophilic archaeon. Thermococcus profundus. The purified protein was shown to be a homomultimer of 93 kDa subunit (P93 complex). The complex is extremely heat stable. During 12 hrs incubation with SDS (final concentration 1%) at $85^{\circ}C$, no changed structure could be observed. Electron image analysis of negatively stained showed that the complex has a single, stable characteristic view and a well-preserved core with threefold rotational symmetry. The periphery of the assembly is composed of a nebulose, possibly flexible, component. Based on the projected structure suggest the P93 complex from T. profundus is composed 24 homomultimer.

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Purification and Biochemical Properties of Storage Protein SP-2 in Tobacco Budworm (Helicoverpa assulta Guenee) (담배나방 저장단백질 SP-2의 정제 및 생화학적 특성)

  • 정성은;채순용;김선봉;이형철
    • Journal of the Korean Society of Tobacco Science
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    • v.18 no.1
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    • pp.39-48
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    • 1996
  • A storage protein(SP-2) was confirmed in haemolymph during larval-pupal-adult development of tobacco burdworm(Helicoverpa assulta Guenee), and its biochemical characteristics were investigated. The titer of SP-2 showed a peak at mature larva, decreased gradually through the late pupal stage, and became undetectable at adult period. As the results from electrophoretic mnysis, SP-2 was confirmed to be glycolipoprotein(M.W. 332kDa) relatively stable to heat( $\leq$ 68$^{\circ}C$ ). This storage protein was determined to be a tetramer composed of a single subunit with MW of 83kDa, and the isoelectric point was 5.7. The amino acid composition of the SP-2 was characterized. It has relatively high content of methionine and histidine, whereas the contents of tyrosine and phenylalanine were relatively low.

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