• Title/Summary/Keyword: p23

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Structural Characterization of Growth-Related Translationally Controlled Tumor Protein P23

  • Lee, Bong-Jin;Hong, Yoon-Hun;Park, Sang-Ho;Lee, Kyunglim
    • Journal of the Korean Magnetic Resonance Society
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    • v.5 no.1
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    • pp.46-55
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    • 2001
  • P23, a translationally controlled turner protein is involved in the interleukin-4 secretion from human basophils and is also known to be an IgE-dependent histamine-releasing factor. However, the precise physiological function and structure of P23 have not been elucidated. In the current study, we constructed the optimal expression and purification protocol of P23 and investigated the secondary structure and structural stability in various conditions. Circular dichroism (CD) investigation showed that the secondary structure of P23 adopts mainly a P-sheet conformation. CD spectroscopy and differential scanning calorimetry revealed that P23 is fairly stable in the pH range of neutral and mild-basic conditions and in the temperature range of 10 - 50$\^{C}$. Since the thermal stability and the P-sheet content of P23 were decreased by the addition of Ca$\^$2+/ ion, it could be suggested that Ca$\^$2+/ion induces structural change by partially destabilizing the structure of P23. In addition various H experiments were monitored to solve the aggregation of P23. Den results will provide the preliminary structural information about P23.

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Study of an ER bound p80 Homologous to Nucleolar B23 (핵소체 단백 B23과 세포질 단백 p80의 유사성에 관한 연구)

  • Lee, Hye-Jeong;Yoon, Sang-In;Choi, Yong-Chun;Ahn, Young-Soo
    • The Korean Journal of Pharmacology
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    • v.31 no.2
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    • pp.241-250
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    • 1995
  • Protein B23 is one of the major nucleolar phosphoproteins associated with pre-ribosomal particles, and is localized in the granular region of the nucleolus. Recent studies suggest that protein B23 shuttles between nucleus and cytoplasm and also interacts with HIV Rev. These findings indicate that protein B23 is important in nucleocytoplasmic relationship and viral replication. However, the exact function of protein B23 is not clear yet. In acute nucleolar hypertrophy of rat liver, treated with thioacetamide, there was observed an increase of not only protein B23 but also B23-like protein p45 when anti-B23 monoclonal antibody (MAb) was used for identification. On the basis of the large B23 specific epitope structure composed of 68 amino acids, a hypothesis was formulated to examine that p45 is the pre-B23 resulting from excessive production of B23. In an attempt to investigate the precursor of B23, we analyzed the subcellular fractions and microsomal subfractions. Subsequently, we analyzed the finger printings of B23-like proteins using the tryptic peptide mapping. The results are summarized: 1) Using B23 MAb, we observed the presence of B23-like proteins in nucleolar fraction, nucleoplasmic fraction and microsomal fraction. 2) In the further microsomal subfractionation, we could partially purify B23-like protein in 2M layer of sucrose gradient. 3) When ion exchange chromatography was employed, there were protein species 80kDa(p80), 65kDa(p65) and 60kDa(p60). 4) Based on the tryptic map analysis of $^{125}I$ labeled proteins, the similarity between B23 and p80 was found only in 9 out of 14(B23) and 21(p80) peptides, and difference was found in the remaining peptides. p80 and p60 had 18 common peptides, and all the peptides of p60 were similar to those of p80. From these results, it is proposed that p45 is an abnormal metabolite resulting from carcinogenesis by thioacetamide, and it is not the precursor of B23. In addition, we suggest that p80 may be a precursor of p45.

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The Stability of p53 in Ras-mediated Senescent Cells in Response to Nucleolar Stress (Ras에 의해 유도된 노화세포에서 핵인 스트레스에 의한 p53 안정화 연구)

  • Sihn, Choong-Ryoul;Park, Gil-Hong;Lee, Kee-Ho;Kim, Sang-Hoon
    • Journal of Life Science
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    • v.19 no.4
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    • pp.436-441
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    • 2009
  • B23/nucleophosmin, a nucleolar protein, translocates into the nucleus from the nucleolus when cells are damaged by extracellular stresses. Recently, it was shown that such translocation of B23/nucleophosmin in normal fibroblasts under stress conditions increases both the stability and activation of the p53 protein by disrupting its interaction with MDM2. Senescent cells have a single large nucleolus and a diminished capacity to induce p53 stability upon exposure to various DNA damaging agents. To investigate the role of B23/nucleophosmin in p53 stability in senescent cells, we established a senescence model system by expressing the ras oncogene in IMR90 cells. The stability of p53 was reduced in these cells in response to nucleolar stress, although the level of B23/nucleophosmin protein was not changed. In addition, p53 did not accumulate in the nucleus and B23/nucleophosmin did not translocate into the nucleoplasm. The binding affinity of B23/nucleophosmin with p53 was reduced in senescent cells, whereas the interaction between MDM2 and p53 was stable. Taken together, the stability of p53 in ras-induced senescent cells may be influenced by the ability of B23/nucleophosmin to interact with p53 in response to nucleolar stress.

Purification and Characterization of 2,3-Dihydroxybiphenyl 1,2- Dioxygenase from Comamonas sp.

  • Lee Na Ri;Kwon Dae Young;Min Kyung Hee
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2001.11a
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    • pp.16-25
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    • 2001
  • A genomic library of biphenyl-degrading strain Comamonas sp. SMN4 was constructed by using the cosmid vector pWE15 and introduced into Escherichia coli. Of 1,000 recombinant clones tested, two clones that expressed 2,3-dihydroxybiphenyl 1,2-dioxygenase activity were found (named pNB 1 and pNB2). From pNB1 clone, subclone pNA210, demonstrated 2,3-dihydroxybiphenyl 1,2-dioxygenase activity, is isolated. 2,3-Dihydroxybiphenyl 1,2-dioxygenase (23DBDO, BphC) is an extradiol-type dioxygenase that involved in third step of biphenyl degradation pathway. The nucleotide sequence of the Comamonas sp. SMN4 gene bphC, which encodes 23DBDO, was cloned into a plasmid pQE30. The His-tagged 23DBDO produced by a recombinant Escherichia coli, SG 13009 (pREP4)(pNPC), and purified with a Ni-nitrilotriacetic acid resin affinity column using the His-bind Qiagen system. The His-tagged 23DBDO construction was active. SDS-PAGE analysis of the purified active 23DBDO gave a single band of 32 kDa; this is in agreement with the size of the bphC coding region. The 23DBDO exhibited maximum activity at pH 9.0. The CD data for the pHs, showed that this enzyme had a typical a-helical folding structures at neutral pHs ranged from pH 4.5 to pH 9.0. This structure maintained up to pH 10.5. However, this high stable folding strucure was converted to unfolded structure in acidic region (pH 2.5) or in high pH (pH 12.0). The result of CD spectra observed with pH effects on 23DBDO activity, suggested that charge transition by pH change have affected change of conformational structure for 23DBDO catalytic reaction. The $K_m$ for 2,3-dihydroxybiphenyl, 3-metylcatechol, 4-methylcatechol and catechol was 11.7 $\mu$M, 24 $\mu$M, 50 mM and 625 $\mu$M.

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IL-23 Inhibits Trophoblast Proliferation, Migration, and EMT via Activating p38 MAPK Signaling Pathway to Promote Recurrent Spontaneous Abortion

  • He, Shan;Ning, Yan;Ma, Fei;Liu, Dayan;Jiang, Shaoyan;Deng, Shaojie
    • Journal of Microbiology and Biotechnology
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    • v.32 no.6
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    • pp.792-799
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    • 2022
  • As a vital problem in reproductive health, recurrent spontaneous abortion (RSA) affects about 1% of women. We performed this study with an aim to explore the molecular mechanism of interleukin-23 (IL-23) and find optimal or effective methods to improve RSA. First, ELISA was applied to evaluate the expressions of IL-23 and its receptor in HTR-8/SVneo cells after IL-23 treatment. CCK-8, TUNEL, wound healing and transwell assays were employed to assess the proliferation, apoptosis, migration and invasion of HTR-8/SVneo cells, respectively. Additionally, the expressions of apoptosis-, migration-, epithelial-mesenchymal transition- (EMT-) and p38 MAPK signaling pathway-related proteins were measured by western blotting. To further investigate the relationship between IL-23 and p38 MAPK signaling pathway, HTR-8/SVneo cells were treated for 1 h with p38 MAPK inhibitor SB239063, followed by a series of cellular experiments on proliferation, apoptosis, migration and invasion, as aforementioned. The results showed that IL-23 and its receptors were greatly elevated in IL-23-treated HTR-8/SVneo cells. Additionally, IL-23 demonstrated suppressive effects on the proliferation, apoptosis, migration, invasion and EMT of IL-23-treated HTR-8/SVneo cells. More importantly, the molecular mechanism of IL-23 was revealed in this study; that is to say, IL-23 inhibited the proliferation, apoptosis, migration, invasion and EMT of IL-23-treated HTR-8/SVneo cells via activating p38 MAPK signaling pathway. In conclusion, IL-23 inhibits trophoblast proliferation, migration, and EMT via activating p38 MAPK signaling pathway, suggesting that IL-23 might be a novel target for the improvement of RSA.

Clinical Significance of p53 Gene and nm23 Gene Expression in Esophageal Cancer (식도암 조직에시 p53 및 nm23 유전자 발현의 임상적 의의)

  • Park, Kuhn;Lee, Jong-Ho;Sa, Young-Jo;Jin, Ung;Kwon, Jong-Bum;Park, Jae-Gil;Lee, Sun-He;Kwak, Moon-Sub
    • Journal of Chest Surgery
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    • v.37 no.3
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    • pp.261-266
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    • 2004
  • Although significant progress has been made in the surgical treatment of esophageal carcinoma as well as in the detection of early stage esophageal carcinoma by diagnostic techniques, the prognosis of the esophageal carcinoma patients remain poor. The p53 gene product is known to regulate cell growth and proliferation. And the nm23 gene was identified originally as an anti-metastatic influence whose expression was correlated inversely with tumor metastatic potential in murine melanoma cell lines. This experiment was intended to know the relationship among the p53 and nm23 gene expression versus clinicopahologic characteristics of the esophageal cancer. Total 40 cases were collected from patients who had undergone esophagectomy at St. Mary's Hospital, Catholic university of Korea. Immunohistochemical stain for p53 mutant-type protein and nm23 protein was graded as <10% positive tumor cells: negative; 10∼30% positive tumor cells: + ; 30∼50% positive tumor cells: ++, and >50% positive tumor cells: +++. The tumor invasion was grades as none:- ; mild:+ ; moderate:++ ; severe: +++. Overexpression of p53 protein and nm23 was not associated with the survival and cliniocopathologic characteristics of the esophageal cancer. Moreover, the combination analysis of p53 and nm23 revealed that there was no relationship between the gene expression and the clinicopatholic characteristics of the esophageal cancer.

6p23 Deletion Syndrome : Report of a Case in a Preterm Baby (미숙아에서의 6p23 Deletion Syndrome 1례)

  • Lee, Hyeon-Soo
    • Clinical and Experimental Pediatrics
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    • v.46 no.1
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    • pp.83-85
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    • 2003
  • Deletion of the short arm of chromosome 6 is relatively rare, with the characteristic features of craniofacial malformations, hypotonia, and defects of the heart and kidney, with hydrocephalus and eye abnormalities. Here author reports a premature girl with bilateral anophthalmia, bilateral hydrocephalus and marked hypotonia, whose chromosome analysis revealed a 46, XX, del(6)(p23) chromosome constitution.

IL-23 P19 Expression Induced by IL-17 and $IL-1{\beta}$ in Rheumatoid Arthritis Synovial Mononuclear Cells (류마티스관절염 환자의 활액 세포에서 IL-17과 $IL-1{\beta}$에 의한 IL-23p19의 발현 증가)

  • Cho, Mi-La;Heo, Yu-Jung;Oh, Hye-Jwa;Kang, Chang-Min;Lee, Seon-Yeong;Hong, Yeon-Sik;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.8 no.1
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    • pp.29-37
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    • 2008
  • Interleukin-23 (IL-23) is a novel pro-inflammatory cytokine which has been implicated to play a pathogenic role in rheumatoid arthritis (RA). This study was undertaken to investigate the IL-23 inductive activity of the proinflammatory cytokine IL-17, $IL-1{\beta}$ and tumor necrosis factor (TNF-${\alpha}$) in RA synovial fluid mononuclear cells (SFMC). Expression of IL-23p19, IL-17, $IL-1{\beta}$ and TNF-${\alpha}$ in joint was examined by immunohistochemistry (IHC) of patients with RA and osteoarthritis (OA). The effects of IL-17 and $IL-1{\beta}$ on expression of IL-23p19 in human SFMC from RA patients were determined by reverse transcriptase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). IL-23p19 was expressed in the RA fibroblast like synoviocyte (FLS), but not from OA FLS. Similar to the protein expression, IL-23p19 mRNA could be detected by RT-PCR in RA SFMC. IL-17 and $IL-1{\beta}$ could induce RA SFMC to produce the IL-23p19. The effects of IL-17 were much stronger than $IL-1{\beta}$ or TNF-${\alpha}$. These responses were observed in a doseresponsive manner. In addition, IL-17 or $IL-1{\beta}$ neutralizing antibody down-regulated the expression of IL-23p19 induced by LPS in RA-SFMC. Our results demonstrate that IL-23p19 is overexpressed in RA synovium and IL-17 and $IL-1{\beta}$ appears to upregulate the expression of IL-23p19 in RA-SFMC.

Differentiation of Major Rice-Seedborne Bacteria by PCR-Amplified Polymorphism of Spacer Region Between 16S and 23S Ribosomal DNA (PCR로 증폭된 16S와 23S rDNA 사이 Spacer 부위의 다형성에 의한 주요 벼종자전염성 세균의 구별)

  • 김형무;송완엽
    • Korean Journal Plant Pathology
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    • v.12 no.1
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    • pp.11-20
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    • 1996
  • 한 쌍의 R16-1과 R23-2R primer를 이용한 PCR에 의해 증폭된 16S와 23S rDNA 사이의 rDNA spacer 부위의 다형성들이 Pseudomonas avenae, P. glumae, P. fuscovaginae, P. syringae pv. syrngae, Xanthomonas oryzae pv. oryzae, X. oryzae, Xanthomonas herbicola 등 벼 종자전염성 51개 균주의 구분을 위하여 적용되었다. 증폭산물은 820∼950bp의 크기였으며, 각각의 종에 특이적이었고 구분이 가능하였다. Pseudomonas species의 증폭산물은 P. avenae는 950bp, P. glumae는 850bp, P. fuscovaginae는 770pb 및 P. syringae pv. syringae는 1,240, 1,100 및 820bp로 특이적이었다. P. avenae와 P. glumae의 국내균주들은 다형성에 있어 종내 변이는 없었다. X. oryzae pv. oryzae의 860bp와 X. oryzae pv. oryzicola의 890, 440 및 370bp의 이차산물에서 Xanthomonas species의 종내에서 균주에 관련없이 단일화된 다형성을 보였다. CXO 211을 제외한 모든 국내 균주는 a형에 속한 반면 하나의 국내 균주를 포함하여 4개 균주는 b형이었다. E. herbicola의 spacer 부위 증폭은 여러 개의 band를 보였으며, 증폭상은 각각 동일하였고, strain간의 종내 변이는 없었다. 본 실험 결과에 의하여 16S와 23S rDNAdp R16-1과 R23-2R primer를 이용하여 PCR 증폭된 spacer 다형성의 구별은 종자전염성 세균의 신속한 구별에 이용될 수 있을것이다.

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