• 제목/요약/키워드: Functional characterization

검색결과 794건 처리시간 0.057초

Proteomic Functional Characterization of Bovine Stromal Vascular Cells from Omental, Subcutaneous and Intramuscular Adipose Depots

  • Rajesh, Ramanna Valmiki;Kim, Seong-Kon;Park, Mi-Rim;Nam, Jin-Seon;Kim, Nam-Kuk;Kwon, Seulemina;Yoon, Du-Hak;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Asian-Australasian Journal of Animal Sciences
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    • 제24권1호
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    • pp.110-124
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    • 2011
  • Anatomically separate fat depots differ in size, function, and contribution to pathological states such as the metabolic syndrome. We isolated pre-adipocytes from different adipose depots, omental, subcutaneous and intramuscular, of beef cattle, and cultured in vitro to determine the basis for the variations and attribute these variations to the inherent properties of adipocyte progenitors. The proliferating cells from all depots before the confluence were harvested and the proteome was analyzed by a functional proteomic approach, involving 2-DE and MALDI-TOF/TOF. More than 252 protein spots were identified, selected and analyzed by Image Master (ver 7.0) and MALDI-TOF/TOF. Further, our analysis showed that there were specific differences in proteome expression patterns among proliferating precursor cells from the three depots. Sixteen proteins were found to be differentially expressed and these were identified as proteins involved in cellular processes, heat shock/chaperones, redox proteins, cytoskeletal proteins and metabolic enzymes. The results also enabled us to understand the basic roles of these proteins in different inherent properties exhibited by adipose tissue depots.

Comparative Reverse Screening Approach to Identify Potential Anti-neoplastic Targets of Saffron Functional Components and Binding Mode

  • Bhattacharjee, Biplab;Vijayasarathy, Sandhya;Karunakar, Prashantha;Chatterjee, Jhinuk
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권11호
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    • pp.5605-5611
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    • 2012
  • Background: In the last two decades, pioneering research on anti-tumour activity of saffron has shed light on the role of crocetin, picrocrocin and safranal, as broad spectrum anti-neoplastic agents. However, the exact mechanisms have yet to be elucidated. Identification and characterization of the targets of bioactive constituents will play an imperative role in demystifying the complex anti-neoplastic machinery. Methods: In the quest of potential target identification, a dual virtual screening approach utilizing two inverse screening systems, one predicated on idTarget and the other on PharmMapper was here employed. A set of target proteins associated with multiple forms of cancer and ranked by Fit Score and Binding energy were obtained from the two independent inverse screening platforms. The validity of the results was checked by meticulously analyzing the post-docking binding pose of the picrocrocin with Hsp90 alpha in AutoDock. Results: The docking pose reveals that electrostatic and hydrogen bonds play the key role in inter-molecular interactions in ligand binding. Picrocrocin binds to the Hsp90 alpha with a definite orientation appropriate for nucleophilic attacks by several electrical residues inside the Hsp90-alpha ATPase catalytic site. Conclusion: This study reveals functional information about the anti-tumor mechanism of saffron bioactive constituents. Also, a tractable set of anti-neoplastic targets for saffron has been generated in this study which can be further authenticated by in vivo and in vitro experiments.

프로페닐 에테르 단량체들의 합성과 광중합 특성 (Synthesis and Photopolymerization Characterization of Propenyl Ether Monomers)

  • 김기상;심상연
    • 한국응용과학기술학회지
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    • 제34권2호
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    • pp.203-209
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    • 2017
  • 양이온계 광중합에 적용 가능한 propenyl ether 형태의 단량체들을 mono 및 di-functional alcohol과 allyl bromide의 축합반응으로 합성하였다. 이들 단량체들을 양이온 광개시제와 혼용하여 코팅조성물을 제조하여 광경화 반응성을 조사하였다. 그 결과 mono propenyl ether 형태인((prop-1-en-1-yloxy)methyl)benzene (POMB)는 dipropenyl ether 계인 1,4-bis(prop-1-en-1-yloxy) benzene (BPOB) 비교하여 초기 중합 속도는 10.2로 상대적으로 낮게 나타났으나 1.5mol%의 광개시제를 포합한 배합물에서는 90초이내에 거의 정량적으로 반응하였다. 또한 긴알킬기를 갖는 술폰산 염계 광개시제들은 단량체들에 빠른 용해 특성을 나타내었고 산성도가 높은 광개시제인(4-n-decyloxyphenyl)diphenylsulfonium hexafluoroantimonate(DPSA)와 (4-n-decyloxyphenyl) diphenylsulfonium triflate (DPST)를 사용한 경우 비교적 높은 중합속도와 전환율을 나타내었다.

아크릴 수지로 코팅한 배수성 아스팔트 콘크리트의 특성 평가 (Characterization of Porous Asphalt Concrete Coated with MMA Resin)

  • 최태준;이현종;김태우;송재혁
    • 한국도로학회논문집
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    • 제11권2호
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    • pp.111-120
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    • 2009
  • 본 연구는 배수성 아스팔트 포장의 표면을 아크릴 수지로 코팅 처리할 경우 내구성 측면 및 기능적 측면에서 어떠한 효과가 있는가를 실내시험을 통해 평가하는 기초적인 연구이다. 포장의 내구성 측면에서는 칸타브로 및 휠트래킹, 수분손상 간접 인장 피로 시험 등을 실시하였다. 시험결과 시편을 수지로 코팅을 한 경우 칸타브로 손실률이 3배 정도 감소되었고 균열에 대한 저항성도 대폭 향상되었으나 수분손상 및 소성변형의 저항성은 큰 차이가 없는 것으로 나타났다. 기능적인 측면에서는 코팅 전 후의 투수계수 및 공극률 BPT(British Pendulum Tester)를 사용한 미끄럼 저항성의 변화를 측정하였다. 시험결과 공극률 및 투수계수는 약간 저하되나 기능상의 문제는 없을 것으로 판단되며, 미끄럼 저항성은 규사를 살포하지 않을 경우 미끄럼 저항성이 감소되나 규사를 살포할 경우 코팅 전 수준 이상의 미끄럼 저항성을 확보하는 것으로 나타났다.

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용존산소농도와 pH의 동시 검출용 이중층 광학 센서막의 특성 및 발효공정의 온라인 모니터링에의 응용 (Characterization of Double-layer Optical Sensing Membranes for Dual Sensing of Dissolved Oxygen Concentrations and pH and Their Application to the On-line Monitoring of Fermentation Processes)

  • 김춘광;이종일
    • 공업화학
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    • 제20권4호
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    • pp.370-374
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    • 2009
  • 용존산소농도(DO)와 pH를 동시에 검출하기 위하여 이중층 광학 센서 막을 제조하였다. DO 민감성 염료인 Rudpp를 MTMS 졸-겔에 혼합하고 24-웰 마이크로타이터 플레이트에 코팅하였다. DO-검출층 위에 HPTS와 혼합된 GA 졸-겔용액을 코팅하고 pH 측정을 위해 사용하였다. 이중층 광학 센서 막은 온도와 이온 강도에 영향을 받았다. 또한 DO와 pH 이중층 광학 센서 막은 미생물발효공정에 온라인 모니터링 하는데 응용하였으며 좋은 성능을 보였다.

Functional Characterization and Application of the HpOCH2 Gene, Encoding an Initiating $\alpha$l,6-Mannosyltransferase, for N-glycan Engineering in the Methylotrophic Yeast Hansenula polymorpha

  • Kim, Moo-Woong;Kim, Eun-Jung;Kim, Jeong-Yoon;Rhee, Sang-Ki;Kang, Hyun-Ah
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 2004년도 Annual Meeting BioExibition International Symposium
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    • pp.278-281
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    • 2004
  • The $\alpha$1,6-mannosyltransferase encoded by Saccharomyces cerevisiae OCH1 plays a key role for the outer chain initiation of the N-linked oligosaccharides. A search for Hansenula polymorpha genes homologous to S. cerevisiae OCHI (ScOCH1) has revealed seven open reading frames (ORF100, ORF142, ORF168, ORF288, ORF379, ORF576, ORF580). All of the seven ORFs are predicted to be a type II integral membrane protein containing a transmembrane domain near the amino-terminal region and has a DXD motif, which has been found in the active site of many glycosyltransferases. Among this seven-membered OCH1 gene family of H. polymorpha, we have carried out a functional analysis of H. polymorpha ORF168 (HpOCH2) showing the highest identity to ScOCH1. Inactivation of this protein by disruption of corresponding gene resulted in several phenotypes suggestive of cell wall defects, including hypersensitivity to hygromycin B and sodium deoxycholate. The structural analysis of N-glycans synthesized in HpOCH2-disrupted strain (Hpoch2Δ) and the in vitro $\alpha$1,6-mannosyltransferase activity assay strongly indicate that HpOch2p is a key enzyme adding the first $\alpha$1,6-mannose residue on the core glycan Man$_{8}$GlcNAc$_2$. The Hpoch2Δ was further genetically engineered to synthesize a recombinant glycoprotein with the human compatible N-linked oligosaccharide, Man$_{5}$GlcNAc$_2$, by overexpression of the Aspergillus saitoi $\alpha$1,2-mannosidase with the 'HDEL” ER retention signal.gnal.

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식품, 신선편이 농산물용 저장기간 연장 포장 소재 특성 및 평가 (Characterization of Shelf Life Extension Packaging Material for Food and Fresh Cut Agricultural Product: A Review)

  • 이진규;유지예;김미경;유영선
    • 한국포장학회지
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    • 제22권3호
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    • pp.119-125
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    • 2016
  • 신선편이 농산물은 소비자에게 편이성을 제공하면서 수요가 크게 증가하고 있으나 가공과정 중 절단, 박피 등의 물리적인 변화로 인하여 쉽게 품질이 변하거나 미생물 증식 등이 발생할 수 있다. 따라서 이러한 신선편이 농산물의 저장성 및 안정성 향상을 목적으로 필름이나 용기 형태의 기능성 포장재를 개발하고자 하는 노력이 지속되고 있으며, 아울러 저장기간 연장에 관한 연구도 활발히 진행되고 있다. 이와 관련하여 저장기간에 영향을 주는 항균, 원적외선, 통기성, 방담, 미약전류, 에틸렌가스 흡착 또는 분해, MA (Modified Atmosphere) 또는 CA (Controlled Atmosphere)와 같은 기체 조성 변화 등을 통해 저장기간을 연장시키는 방법이 주목받고 있는데, 한 가지에 의해서가 아니라 여러 가지 복합적인 요인들이 작용하여 저장기간을 연장시키는 방법이 효과적으로 알려져 있다. 따라서 보다 효과적으로 신선편이 농산물, 식품 등의 저장기간을 연장시키기 위해 발표된 문헌자료를 토대로 저장기간 연장 포장법, 저장기간 연장 포장소재, 종합적인 평가 방법 등을 중심으로 정리하였다.

Cloning and Characterization of cDNA Encoding Potentially Functional Mouse Glandular Kallikrein

  • Kim, Hwa-Seon;Kim, Won-Sin
    • BMB Reports
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    • 제30권5호
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    • pp.356-361
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    • 1997
  • We cloned a cDNA (pPRC-1) which was comprised of 841 nucleotides from the cDNA library of a male ICR mouse submandibular gland ($SMG^+$). The nucleotide sequences of pPRC-1 were identical to those of exons 2 and 3 of the mGK-21 gene, a potentially functional glandular kallikrein identified in a Balb/c mouse, except for one nucleotide residue. Although this substitution changes Ile (ATT) in pPRC-1 to Val (GTT) in mGK-21, this difference has been explained by strain polymorphism. From the amino acid sequences predicted from its cDNA, we speculated that mGK-21 gene products/pGK21 consist of 261 amino acids including the $NH_2$-terminal signal peptide (residues 1~17), the short propeptide (residues 17~24), and the active peptide (residues 25~261). Although we did not demonstrate the enzyme activity of pGK21, it was assumed that pGK 21 was involved in the maturation of certain bioactive polypeptide(s) in mouse SMG for the following reasons : (a) mGK-21 gene was apparently expressed in a male ICR mouse SMG: (b) the proposed active site $His^{65}$, $Asp^{120}$, and $Ser^{213}$ residues were completely conserved in pGK21 just like other glandular kallikreins; (c) the cloned cDNA was translated to a predicted 27 kDa polypeptide chain in vitro: (d) the 27 kDa polypeptide chain produced by CHO cells was produced to a putative active form by trypsin.

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Functional Roles of a Putative B' Delta Regulatory Subunit and a Catalytic Subunit of Protein Phosphatase 2A in the Cereal Pathogen Fusarium graminearum

  • Kim, Hee-Kyoung;Yun, Sung-Hwan
    • The Plant Pathology Journal
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    • 제28권3호
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    • pp.259-269
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    • 2012
  • Protein phosphatase 2A (PP2A), a family of serine/threonine protein phosphatases, plays an important role in balancing the phosphorylation status of cellular proteins for regulating diverse biological functions in eukaryotic organisms. Despite intensive studies in mammals, limited information on its role is available in filamentous fungi. Here, we investigated the functional roles of genes for a putative B' delta regulatory subunit (FgPP2AR) and a catalytic subunit (FgPP2AC) of PP2A in a filamentous ascomycete, Fusarium graminearum. Molecular characterization of an insertional mutant of this plant pathogenic fungus allowed us to identify the roles of FgPP2AR. Targeted gene replacement and complementation analyses demonstrated that the deletion of FgPP2AR, which was constitutively expressed in all growth stages, caused drastic changes in hyphal growth, conidia morphology/germination, gene expression for mycotoxin production, sexual development and pathogenicity. In particular, overproduction of aberrant cylindrical-shaped conidia is suggestive of arthroconidial induction in the ${\Delta}FgPP2AR$ strain, which has never been described in F. graminearum. In contrast, the ${\Delta}FgPP2AC$ strain was not significantly different from its wild-type progenitor in conidiation, trichothecene gene expression, and pathogenicity; however, it showed reduced hyphal growth and no perithecial formation. The double-deletion ${\Delta}FgPP2AR;{\Delta}FgPP2AC$ strain had more severe defects than single-deletion strains in all examined phenotypes. Taken together, our results indicate that both the putative regulatory and catalytic subunits of PP2A are involved in various cellular processes for fungal development in F. graminearum.

Proteome Changes in Penicillium expansum Grown in a Medium Derived from Host Plant

  • Xia, Xiaoshuang;Li, Huan;Liu, Fei;Zhang, Ye;Zhang, Qi;Wang, Yun;Li, Peiwu
    • Journal of Microbiology and Biotechnology
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    • 제27권3호
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    • pp.624-632
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    • 2017
  • Penicillium expansum causes blue mold rot, a prevalent postharvest disease of pome fruit, and is also the main producer of the patulin. However, knowledge on the molecular mechanisms involved in this pathogen-host interaction remains largely unknown. In this work, a two-dimensional gel electrophoresis-based proteomic approach was applied to probe changes in P. expansum 3.3703 cultivated in apple juice medium, which was used to mimic the in planta condition. The results showed that the pH value and reducing sugar content in the apple juice medium decreased whereas the patulin content increased with the growing of P. expansum. A total of 28 protein spots that were up-regulated in P. expansum when grown in apple juice medium were identified. Functional categorization revealed that the identified proteins were mainly related to carbohydrate metabolism, secondary metabolism, protein biosynthesis or degradation, and redox homeostasis. Remarkably, several induced proteins, including glucose dehydrogenase, galactose oxidase, and FAD-binding monooxygenase, which might be responsible for the observed medium acidification and patulin production, were also detected. Overall, the experimental results provide a comprehensive interpretation of the physiological and proteomic responses of P. expansum to the host plant environment, and future functional characterization of the identified proteins will deepen our understanding of fungi-host interactions.