• 제목/요약/키워드: PAO

검색결과 122건 처리시간 0.026초

Phenylarsine oxide와 adenosine에 민감한 sulfatide 결합 펩타이드의 trans-golgi network 타기팅 (Phenylarsine Oxide and Adenosine-sensitive Trans-golgi Complex Targeting of GFP Fused to Modified Sulfatide-binding Peptide)

  • 전용우;이진아;장덕진
    • 생명과학회지
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    • 제28권2호
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    • pp.162-169
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    • 2018
  • 세포기질에 존재하는 많은 종류의 단백질들은 N-말단에 존재하는 짧은 펩타이드들에 의해서 trans-golgi network(TGN)의 세포질쪽 막에 타기팅될때 중요한 역할을 수행한다고 보고되고 있다. 본 연구실에서도 이전에 바다달팽이인 군소에서 클로닝된 phosphodiesterase 4의 long-form의 경우 N-말단에 존재하는 20개의 아미노산 서열만으로도 충분히 HEK293T세포의 TGN의 세포질막에 타기팅 되게 하며, 이 펩타이드가 sulfatide와 PI4P에 결합성이 있다는 사실을 in vitro에서 확인하였다. 그래서, 본 연구에서는 sulfatide결합성과 TGN막 타기팅과의 연관성을 연구하고자 하였다. 이를 위해 우선 이전 문헌을 통해 sulfatide결합 펩타이드를 찾았고, 이를 GFP단백질과 융합하여 재조합 단백질(mHSBP-EGFP)을 만들어 세포내 타기팅을 실험해 보았다. 이러한 연구를 수행한 결과, mHSBP-EGFP가 HEK293T세포에서 TGN에 타기팅 되고, sulfatide결합이 망가진 돌연변이는 타기팅이 사라짐을 확인하였다. 또한, mHSBP-EGFP가 TGN에 타기팅 되는 것은 억제제인 antimycin A와 PAO와 adenosine에 의해 억제됨을 확인할 수 있었다. 이러한 사실을 통해, PAO와 adenosine에 민감한 인산화효소들, 그중에 PI4KII의 활성이 mHSBP-EGFP를 TGN으로 위치하게 하는데 중요한 역할을 수행한다고 추론할 수 있다.

Protein phosphorylation on tyrosine restores expression and glycosylation of cyclooxygenase-2 by 2-deoxy-D-glucose-caused endoplasmic reticulum stress in rabbit articular chondrocyte

  • Yu, Seon-Mi;Kim, Song-Ja
    • BMB Reports
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    • 제45권5호
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    • pp.317-322
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    • 2012
  • 2-deoxy-D-glucose(2DG)-caused endoplasmic reticulum (ER) stress inhibits protein phosphorylation at tyrosine residues. However, the accurate regulatory mechanisms, which determine the inflammatory response of chondrocytes to ER stress via protein tyrosine phosphorylation, have not been systematically evaluated. Thus, in this study, we examined whether protein phosphorylation at tyrosine residues can modulate the expression and glycosylation of COX-2, which is reduced by 2DG-induced ER stress. We observed that protein tyrosine phosphatase (PTP) inhibitors, sodium orthovanadate (SOV), and phenylarsine oxide (PAO) significantly decreased expression of ER stress inducible proteins, glucose-regulated protein 94 (GRP94), and CCAAT/ enhancer-binding-protein- related gene (GADD153), which was induced by 2DG. In addition, we demonstrated that SOV and PAO noticeably restored the expression and glycosylation of COX-2 after treatment with 2DG. These results suggest that protein phosphorylation of tyrosine residues plays an important role in the regulation of expression and glycosylation during 2DG-induced ER stress in rabbit articular chondrocytes.

Polyamines and Their Metabolites as Diagnostic Markers of Human Diseases

  • Park, Myung Hee;Igarashi, Kazuei
    • Biomolecules & Therapeutics
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    • 제21권1호
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    • pp.1-9
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    • 2013
  • Polyamines, putrescine, spermidine and spermine, are ubiquitous in living cells and are essential for eukaryotic cell growth. These polycations interact with negatively charged molecules such as DNA, RNA, acidic proteins and phospholipids and modulate various cellular functions including macromolecular synthesis. Dysregulation of the polyamine pathway leads to pathological conditions including cancer, inflammation, stroke, renal failure and diabetes. Increase in polyamines and polyamine synthesis enzymes is often associated with tumor growth, and urinary and plasma contents of polyamines and their metabolites have been investigated as diagnostic markers for cancers. Of these, diacetylated derivatives of spermidine and spermine are elevated in the urine of cancer patients and present potential markers for early detection. Enhanced catabolism of cellular polyamines by polyamine oxidases (PAO), spermine oxidase (SMO) or acetylpolyamine oxidase (AcPAO), increases cellular oxidative stress and generates hydrogen peroxide and a reactive toxic metabolite, acrolein, which covalently incorporates into lysine residues of cellular proteins. Levels of protein-conjuagated acrolein (PC-Acro) and polyamine oxidizing enzymes were increased in the locus of brain infarction and in plasma in a mouse model of stroke and also in the plasma of stroke patients. When the combined measurements of PC-Acro, interleukin 6 (IL-6), and C-reactive protein (CRP) were evaluated, even silent brain infarction (SBI) was detected with high sensitivity and specificity. Considering that there are no reliable biochemical markers for early stage of stroke, PC-Acro and PAOs present promising markers. Thus the polyamine metabolites in plasma or urine provide useful tools in early diagnosis of cancer and stroke.

Effects of various prophylactic procedures on titanium surfaces and biofilm formation

  • Di Salle, Anna;Spagnuolo, Gianrico;Conte, Raffaele;Procino, Alfredo;Peluso, Gianfranco;Rengo, Carlo
    • Journal of Periodontal and Implant Science
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    • 제48권6호
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    • pp.373-382
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    • 2018
  • Purpose: The aim of this study was to evaluate the effects of various prophylactic treatments of titanium implants on bacterial biofilm formation, correlating surface modifications with the biofilms produced by Pseudomonas aeruginosa PAO1, Staphylococcus aureus, and bacteria isolated from saliva. Methods: Pure titanium disks were treated with various prophylactic procedures, and atomic force microscopy (AFM) was used to determine the degree to which surface roughness was modified. To evaluate antibiofilm activity, we used P. aeruginosa PAO1, S. aureus, and saliva-isolated Streptococcus spp., Bacteroides fragilis, and Staphylococcus epidermidis. Results: AFM showed that the surface roughness increased after using the air-polishing device and ultrasonic scaler, while a significant reduction was observed after using a curette or polishing with Detartrine ZTM (DZ) abrasive paste. In addition, we only observed a significant (P<0.01) reduction in biofilm formation on the DZ-treated implant surfaces. Conclusion: In this study, both AFM and antibiofilm analyses indicated that using DZ abrasive paste could be considered as the prophylactic procedure of choice for managing peri-implant lesions and for therapy-resistant cases of periodontitis.

Reg. Guide 1.52(Rev.3)를 적용한 원전 ESF 공기정화계통 성능시험법 개선 연구 (A Study on Improvement of Test Method of Nuclear Power Plant ESF ACS by applying Regulatory Guide 1.52 (Rev.3))

  • 이숙경;김광신;손순환;송규민;이계우;박정서;조병호;유병재;홍순준;강선행
    • 방사성폐기물학회지
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    • 제8권4호
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    • pp.311-318
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    • 2010
  • 공학적 안전설비 공기정화계통의 규제지침인 Reg. Guide 1.52(Rev.3)의 변경사항중 성능시험과 관련한 운전가능성 시험시간 단축, HEPA 필터 현장누설시험용 시험물질 변경 및 활성탄 성능시험 Methyl Iodidie 투과허용율 상향 변경을 영광 5,6호기에 적용하고자, 모사실험장치와 현장 설비를 활용하여 기술적 타당성을 확인하는 실험을 수행하였다. 10시간 이상의 장시간 운전가능성 시험을 통해 계통내 습분을 제거하여도 시험후 1~4일만에 회복됨을 확인하여 운전가능성 시험은 기기적 운전가능성 점검에 적합한 매월 15분 이상의 시험을 수행하는 것이 타당함을 확인하였다. HEPA 필터 현장누설시험용 시험물질 변경을 위해 DOP와 PAO의 에어로졸 입자크기, 발생량, 누설인지도를 비교한 결과 PAO는 원전에서도 DOP 대체시험물질로 사용 가능함을 확인하였다. 베드깊이 4 인치 이상의 활성탄여과기에 대한 Methyl Iodide의 투과율 허용치가 0.175 %에서 0.5 %로 상향 변경된 것은, ASTM D3803(1989)으로의 활성탄 성능시험 방법 변경에 따른 것으로서, $30^{\circ}C$ 상대습도 95 %에서의 Methyl Iodide 투과허용율 0.5 %가 사용 중 활성탄의 성능을 시험하기에 충분히 보수적인 시험방법임을 확인하였다. 본 실험 결과를 바탕으로 영광 5,6호기는 인허가변경을 완료하였다.

Inhibition of Microbial Quorum Sensing Mediated Virulence Factors by Pestalotiopsis sydowiana

  • Parasuraman, Paramanantham;Devadatha, B;Sarma, V. Venkateswara;Ranganathan, Sampathkumar;Ampasala, Dinakara Rao;Reddy, Dhanasekhar;Kumavath, Ranjith;Kim, In-Won;Patel, Sanjay K.S.;Kalia, Vipin Chandra;Lee, Jung-Kul;Siddhardha, Busi
    • Journal of Microbiology and Biotechnology
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    • 제30권4호
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    • pp.571-582
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    • 2020
  • Quorum sensing (QS)-mediated infections cause severe diseases in human beings. The control of infectious diseases by inhibiting QS using antipathogenic drugs is a promising approach as antibiotics are proving inefficient in treating these diseases. Marine fungal (Pestalotiopsis sydowiana PPR) extract was found to possess effective antipathogenic characteristics. The minimum inhibitory concentration (MIC) of the fungal extract against test pathogen Pseudomonas aeruginosa PAO1 was 1,000 ㎍/ml. Sub-MIC concentrations (250 and 500 ㎍/ml) of fungal extract reduced QS-regulated virulence phenotypes such as the production of pyocyanin, chitinase, protease, elastase, and staphylolytic activity in P. aeruginosa PAO1 by 84.15%, 73.15%, 67.37%, 62.37%, and 33.65%, respectively. Moreover, it also reduced the production of exopolysaccharides (74.99%), rhamnolipids (68.01%), and alginate (54.98%), and inhibited the biofilm formation of the bacteria by 90.54%. In silico analysis revealed that the metabolite of P. sydowiana PPR binds to the bacterial QS receptor proteins (LasR and RhlR) similar to their respective natural signaling molecules. Cyclo(-Leu-Pro) (CLP) and 4-Hydroxyphenylacetamide (4-HPA) were identified as potent bioactive compounds among the metabolites of P. sydowiana PPR using in silico approaches. The MIC values of CLP and 4-HPA against P. aeruginosa PAO1 were determined as 250 and 125 ㎍/ml, respectively. All the antivirulence assays were conducted at sub-MIC concentrations of CLP (125 ㎍/ml) and 4-HPA (62.5 ㎍/ml), which resulted in marked reduction in all the investigated virulence factors. This was further supported by gene expression studies. The findings suggest that the metabolites of P. sydowiana PPR can be employed as promising QS inhibitors that target pathogenic bacteria.