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구인 peroxidase의 부분 정제 및 효소학적 특성 규명
이미영,김윤경,이기완 순천향대학교 기초과학연구소 1997 순천향자연과학연구 논문집 Vol.3 No.2
Peroxidase has been partially purified from earthworm, Lumbricus rubellus, using ammonium sulfate fractionation, Sephacryl S-200 gel filtration, CM-cellulose cation exchange chromatography and native-PAGE elution. Some of its enzymatic characteristics were examined. Earthworm peroxidase showed the characteristics of heat-sensitivity, and the enzyme activity was stimulated in the presence of various cations. The most available booster was CoCl₂, followed by ZnCl₂and KCl among cations used. The optimum pH for guaiacol oxidation of earthworm peroxidase was determined to be 6.0, and the Km values against guaiacol and H₂O₂were 1.25mM and 3.4 mM, respectively. When various compounds were tested as the possible substrates of the enzyme, o-dianisidine was used as the substrate. However, earthworm peroxidase could not oxidize esculetin and ferulic acid as substrates, suggesting the different characteristics of the enzyme from plant peroxidases. The opimum pH for veratryl alcohol and H₂O₂oxidation was determined to be 2.5 when lignin peroxidation activity was examined. The Km values for veratryl alcohol and H₂O₂were 0.02 mM and 0.13 mM, respectively.
RAW264.7 세포에서 interferon-r 및 LPS에 의해 유도되는 NO생성에 미치는 TALT-35의 영향
박종일,박경석,김종석,박지훈,윤은진,송경섭,서강식,김훈,윤완희,박승길,임규,황병두 충남대학교 생물공학연구소 2006 생물공학연구지 Vol.12 No.-
TALP-35 purified from human term placenta is known to increase microtubule polymerization and stabilize the polymerized microtubule. To examine the effect of TALP-35 on immune system this study was performed. MTT assay was performed to investigate the effect of TALP-35 on the proliferation of RAW264.7 cells. TALP-35 dose dependently suppress the proliferation of RAW264.7 cells at high concentration (above 1 μM) in unstimulated cells, in case of 10 μM TALP-35 treated cells the suppression was 25% but in stimulated cells it was only 15%. Cosedimentation assay was carried out to investigate whether TALP-35 can bind to tubulin of RAW264.7, monocyte/macrophage lineage of mouse, and polymerize it. TALP-35 polymerize the tubulin of RAW264.7 cells and sedimented in dose-dependent manner. To investigate the effect of TALP-35 on the expression of iNOS protein western blotting was performed. The expression level of iNOS was decreased dose dependently in high concentration of TALP-35 treatment. To examine the activity of iNOS, secreted NO was determined by method based on Griess reaction. Interferon-γ and LPS-stimulated production of NO from RAW264.7 cells was decreased dose dependently above 0.1 μM concentration of TALP-35 and 50% is decreased at 10μM of it. This study shows TALP-35 can control cytokine induced-iNOS expression therefore it might control inflammatory diseases.
CIB1 functions as a Ca(2+)-sensitive modulator of stress-induced signaling by targeting ASK1.
Yoon, Kyoung Wan,Cho, Jun-Ho,Lee, Jae Keun,Kang, Young-Hee,Chae, Ji Soo,Kim, Young Mok,Kim, Jeehyun,Kim, Eun Kyung,Kim, Sung Eun,Baik, Ja-Hyun,Naik, Ulhas P,Cho, Ssang-Goo,Choi, Eui-Ju National Academy of Sciences 2009 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF Vol.106 No.41
<P>Calcium and integrin binding protein 1 (CIB1) is a Ca(2+)-binding protein of 22 kDa that was initially identified as a protein that interacts with integrin alpha(IIb). Although it interacts with various proteins and has been implicated in diverse cellular functions, the molecular mechanism by which CIB1 regulates intracellular signaling networks has remained unclear. We now show that, by targeting apoptosis signal-regulating kinase 1 (ASK1), CIB1 negatively regulates stress-activated MAPK signaling pathways. CIB1 was thus shown to bind to ASK1, to interfere with the recruitment of TRAF2 to ASK1, and to inhibit the autophosphorylation of ASK1 on threonine-838, thereby blocking ASK1 activation. Furthermore, CIB1 mitigated apoptotic cell death initiated either by TNF-alpha in breast cancer MCF7 cells or by 6-hydroxydopamine (6-OHDA) in dopaminergic cells. Ca(2+) influx induced by membrane depolarization reversed the inhibitory effect of CIB1 on 6-OHDA-induced ASK1 activation and cell death in dopaminergic neurons. These observations thus suggest that CIB1 functions as a Ca(2+)-sensitive negative regulator of ASK1-mediated signaling events.</P>
Dead cell phagocytosis and innate immune checkpoint
( Kyoung Wan Yoon ) 생화학분자생물학회(구 한국생화학분자생물학회) 2017 BMB Reports Vol.50 No.10
The human body loses several billions of cells daily. When cells die in vivo, the corpse of each dead cell is immediately cleared. Specifically, dead cells are efficiently recognized and cleared by multiple types of neighboring phagocytes. Early research on cell death focused more on molecular mechanisms of cell death regulation while the cellular corpses were merely considered cellular debris. However, it has come to light that various biological stimuli following cell death are important for immune regulation. Clearance of normal dead cells occurs silently in immune tolerance. Exogenous or mutated antigens of malignant or infected cells can initiate adaptive immunity, thereby inducing immunogenicity by adjuvant signals. Several pathogens and cancer cells have strategies to limit the adjuvant signals and escape immune surveillance. In this review, we present an overview of the mechanisms of dead cell clearance and its immune regulations. [BMB Reports 2017; 50(10): 496-503]
Apoptotic cell clearance and human diseases
Yoon, Kyoung Wan Cellmed Orthocellular Medicine and Pharmaceutical 2017 셀메드 (CellMed) Vol.7 No.1
The efficient removal of dead cells is an evolutionarily conserved process essential for homeostasis in multicellular organisms. The phagocytosis involves a series of steps that ultimately leads the detection of apoptotic cell by the phagocytes and the subsequent engulfment and degradation of corpse. The uptake of apoptotic cells by phagocytes not only removes debris from tissues but also generates an anti-inflammatory signal that blocks tissue inflammation. Conversely, impaired clearance of dead cells can cause loss of immune tolerance and the development of various inflammation-associated diseases such as autoimmunity, but can also affect cancer development. This review will discuss current understanding of the molecular mechanism of apoptotic cell phagocytosis and how they may be related to human diseases.
The irony benign tumor of Aggressive angiomyxoma
( Wan-ju Kim ),( Hee-jung Lee ),( Yeon-ji Lee ),( Yoon-kyoung Lee ),( Jin-seok Suck ),( In-ae Cho ),( Jeong-kyu Shin ),( Won-jun Choi ),( Soon-ae Lee ),( Jong-hak Lee ),( Won-young Paik ) 대한산부인과학회 2016 대한산부인과학회 학술대회 Vol.102 No.-
The aggressive angiomyxoma is known for benign tumor because of non-metastasizing and not invading other organs, but it have high recurrence rate. Why it called as aggressive. A 23 year-old-woman who suffered from labial mass came outpatient clinic. Her chief complaint was palpable vulva mass. She had no underlying diseases until knew about it. She was no pain and redness. Her first impression was Bartholin's cyst and we prepared Marsupialization. Used local anesthesia by Lidocain, surgical exision was done. Surprisingly, the result of biopsy was suspicious aggressive angiomyxoma. There have tendency of local recurrence more often. Regrettably, there was no proven treatment of medicine. Some people have tried to effort treatment by chemotherapy. Consequently, the patient need to have a periodic checkup for recurrence the benign myxoid tumor.