• 제목/요약/키워드: protein cross-linking

검색결과 80건 처리시간 0.025초

사람의 타액선에서 proline-rich protein의 발현양상과 기능적 단백 구조에 대한 분자생물학적 연구 (A MOLECULAR BIOLOGICAL STUDY ON THE EXPRESSION PATTERN AND FUNCTIONAL PROTEIN STRUCTURES OF PROLINE-RICH PROTEINS IN HUMAN SALIVARY GLANDS)

  • 주재용;이석근;박영욱
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제28권1호
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    • pp.31-41
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    • 2002
  • Proline-rich proteins (PRPs) are major components of human saliva. In order to know the biological roles of PRPs, we explored the expression pattern and functional protein structures of PRPs by the immunohistochemical and various molecular biological methods. Polyclonal antibody against human gPRP was generated from rabbit by the injection of oral exfoliated cells specially treated by urea and SDS buffer. The PRPs began to be expressed both in the acinar cells and ductal cells from the EIDS (Early Intermediate Developmental Stage) of fetal salivary glands and became intense in the salivary epithelium in the LDS (Late Developmental Stage) and adult salivary glands. The polyclonal antibody against the gPRP showed the cross-reactivity with aPRP and bPRP, these results were relevant to the high homology among subtypes of PRP. However, the simulated protein structures of PRPs showed the characteristic repetitive whorling domains except the N-terminal signal peptide. The whorling domains were also contained the multiple amino acids of glutamine and glycine, which may provide the receptor binding or cross-linking sites of PRPs.

Development of piezoelectric immunosensor for the rapid detection of marine derived pathogenic bacteria, Vibrio vulnificus

  • Hong, Suhee;Jeong, Hyun-Do
    • 한국어병학회지
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    • 제27권2호
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    • pp.99-105
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    • 2014
  • Biosensors consist of biochemical recognition agents like antibodies immobilized on the surfaces of transducers that change the recognition into a measurable electronic signal. Here we report a piezoelectric immunosensor made to detect Vibrio vulnificus. A 9MHz AT-cut piezoelectric wafer attached with two gold electrodes of 5mm diameter was used as the transducer of the QCM biosensor with a reproducibility of ${\pm}0.1Hz$ in frequency response. We have tried different approaches to immobilize antibody on the sensor chip. Concerning the orientation of antibody for the best antigen binding capacity, the antibody was immobilized by specific binding to protein G or by cross-linking through hydrazine. In addition, protein G was cross-linked on glutaraldehyde activated immine layer (PEI) or EDC/NHS activated sulfide monolayer (MPA). PEI was found to be more effective to immobilize protein G following glutaraldehyde activation than MPA. However, hydrazine chip showed a better capability to immobilize more IgG than protein G chip and a higher sensitivity. The sensor system was able to detect V. vulnificus in dose dependent manner and was able to detect bacterial cells within 5 minutes by monitoring frequency shifts in real time. The detection limit can be improved by preincubation to enrich the bacterial cell number.

단백질 glycation 저해효과가 있는 식품소재 (Anti-glycation Activities from Various Agricultural Products)

  • 최희돈;최인욱;김윤숙
    • 한국식품과학회지
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    • 제39권4호
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    • pp.458-463
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    • 2007
  • 식품소재의 단백질 glycation 저해효과를 조사하기 위하여 곡류, 채소류 및 생약재 등 83종 에탄올 추출물에 대한 단백질 가교결합 저해효과를 $[^{14}C]$-N-formyl-lysine incorporation method를 이용하여 측정하였다. 대부분의 추출물에서 저해효과가 나타났으나 일부 추출물은 단백질의 가교를 촉진하는 것으로 나타났다. 상대적으로 높은 활성을 나타낸 시료 중 20개를 선별하여 형광광도법을 이용한 AGE형성 억제능을 조사한 결과, 1일 발아한 메밀의 추출물 (GB-l)이 가장 높은 저해활성을 가지는 것으로 나타났다. 차후 GB-0l은 콜로로포름, 에틸아세테이트, 부탄올 및 물로 분획되었으며 그 중 에틸아세테이트 분획이 가장 높은 단백질 glycation 저해활성(95.2% inhibitory activity at 100 ug/mL addi- tion level)을 보였다. HPLC를 이용한 폴리페놀 분석결과 발아 및 추출, 분획과정을 거친 메밀의 아세테이트 분획은 rutin 및 quercetin 함량이 상당히 증가한 것으로 나타나서 부분정제물의 rutin 및 quercetin 함량과 저해활성사이에 높은 상관성을 나타내었다.

유청단백질을 이용한 미세캡슐의 응용 (Water-insoluble, Whey Protein-based Microcapsules for Controlled Core Release Application)

  • 이성제
    • 한국유가공학회:학술대회논문집
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    • 한국유가공기술과학회 2005년도 창립 30주년 기념 국제심포지움 - 웰빙시대의 우유.유제품의 새로운 발견
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    • pp.37-61
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    • 2005
  • 미세캡슐 (microencapsulation)은 내부 물질(core material)의 방출을 제어하기 위하여 여러 천연 및 생분해성 물질이 사용된다. 유청단백질은 이러한 목적에 아주 적합한 것으로 여겨지고 있는데 이는 독특한 이화학적 특성에 기인되는 것이다. 본 연구팀은 drug이나 생리활성물질을 피복하기 위하여 cross-linking 물질로 glutaraldehyde를 사용하여 수용성 이면서 유청단백질을 근간으로 하는 미세캡슐 제조기술을 개발하였다. 또한 생리적 조건에서 이들 캡슐의 분해 및 포집물질의 방출에 대한 연구를 수행하였는데 수용성 drug으로 사용된 theophylline은 유청단백질에 잘 분산되는 것으로 확인되었다. 이 분산액은 !%의 생리활성물질인 polyurethane을 함유하는 dichloromethane과 hexane 혼합불에 잘 확산이 되었다. 미세캡슐공정에서 피복물질로 사용되는 여러 물질중에서 유청단백질은 생리적 효능이 뛰어나고 여러 물리적 작용이 있기 때문에 새로운 피복소재로써 그 효용성이 매우 높다. 지금까지의 연구결과 유청단백질을 이용한 미세캡슐의 제조는 유청단백질이 가지는 기능적 효과와 내부물질이 지니는 약리효과를 동시에 이용할 수 있다는 점에서 향후 고부가 식품 의약품 첨가물질로써 유용성이 기대된다고 하겠다.

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산업용 효소로써 티로시나아제 연구의 최근 동향 (Recent Advances in Tyrosinase Research as An Industrial Enzyme)

  • 김혜린;김현미;최유성
    • KSBB Journal
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    • 제29권1호
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    • pp.1-8
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    • 2014
  • Tyrosinases catalyze the hydroxylation of monophenolic compounds and the conversion of o-diphenols to oquinones. The enzymes are mainly involved in the modification of tyrosine into L-3,4-dihydroxyphenyl-alanine (L-DOPA) and DOPA/DOPAquinone-drived intermolecular cross-linking, which play the key roles of pigmentation to the cells. It is ubiquitously distributed in microorganisms, plants, and animals all around the nature world. They are classified as copper- containing dioxygen activating enzymes; two copper ions are coordinated with six histidine residues in their active sites and they are distinguished as met-, deoxy-, and oxy-form depending on their oxidative states. Natural extraction and recombinant protein approaches have been tried to obtain practical amounts of the enzymes for industrial application. Tyrosinases have been widely applied to industrial and biomedical usages such as detoxification of waste water containing phenolic compounds, L-DOPA as a drug of Parkinson's disease, biomaterials preparation based on the cross-linking ability and biosensors for the detection of phenolic compounds. Therefore, this review reports the mechanism of tyrosinase, biochemical and structural features and potential applications in industrial field.

Water-insoluble, Whey Protein-based Microcapsules for Controlled Core Release Application

  • Lee, Sung-Je
    • Journal of Dairy Science and Biotechnology
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    • 제23권2호
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    • pp.115-123
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    • 2005
  • Microcapsules consisting of natural, biodegradable polymers for controlled and/or sustained core release applications are needed. Physicochemical properties of whey proteins suggest that they may be suitable wall materials in developing such microcapsules. The objectives of the research were to develop water-insoluble, whey protein-based microcapsules containing a model water-soluble drug using a chemical cross-linking agent, glutaraldehyde, and to investigate core release from these capsules at simulated physiological conditions. A model water soluble drug, theophylline, was suspended in whey protein isolate (WPI) solution. The suspension was dispersed in a mixture of dichloromethane and hexane containing 1% biomedical polyurethane. Protein matrices were cross-linked with 7.5-30 ml of glutaraldehyde-saturated toluene (GAST) for 1-3 hr. Microcapsules were harvested, washed, dried and analyzed for core retention, microstructure, and core release in enzyme-free simulated gastric fluid (SGF) and simulated intestinal fluid(SIF) at $37^{\circ}C$. A method consisting of double emulsification and heat gelation was also developed to prepare water-insoluble, whey protein-based microcapsules containing anhydrous milkfat (AMF) as a model apolar core. AMF was emulsified into WPI solution (15${\sim}$30%, pH 4.5-7.2) at a proportion of 25${\sim}$50%(w/w, on dry basis). The oil-in-water emulsion was then added and dispersed into corn oil ($50^{\circ}C$) to form an O/W/O double emulsion and then heated at $85^{\circ}C$ for 20 min for gelation of whey protein wall matrix. Effects of emulsion composition and pH on core retention, microstructure, and water-solubility of microcapsules were determined. Overall results suggest that whey proteins can be used in developing microcapsules for controlled and sustained core release applications.

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New insight into transglutaminase 2 and link to neurodegenerative diseases

  • Min, Boram;Chung, Kwang Chul
    • BMB Reports
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    • 제51권1호
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    • pp.5-13
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    • 2018
  • Formation of toxic protein aggregates is a common feature and mainly contributes to the pathogenesis of neurodegenerative diseases (NDDs), which include amyotrophic lateral sclerosis (ALS), Alzheimer's, Parkinson's, Huntington's, and prion diseases. The transglutaminase 2 (TG2) gene encodes a multifunctional enzyme, displaying four types of activity, such as transamidation, GTPase, protein disulfide isomerase, and protein kinase activities. Many studies demonstrated that the calcium-dependent transamidation activity of TG2 affects the formation of insoluble and toxic amyloid aggregates that mainly consisted of NDD-related proteins. So far, many important and NDD-related substrates of TG2 have been identified, including $amlyoid-{\beta}$, tau, ${\alpha}-synuclein$, mutant huntingtin, and ALS-linked trans-activation response (TAR) DNA-binding protein 43. Recently, the formation of toxic inclusions mediated by several TG2 substrates were efficiently inhibited by TG2 inhibitors. Therefore, the development of highly specific TG2 inhibitors would be an important tool in alleviating the progression of TG2-related brain disorders. In this review, the authors discuss recent advances in TG2 biochemistry, several mechanisms of molecular regulation and pleotropic signaling functions, and the presumed role of TG2 in the progression of many NDDs.

Finding and Characterization of Viral Nonstructural Small Protein in Prospect Hill Virus Infected Cell

  • 남기연;정동훈;최재원;이윤성;이평우
    • 대한바이러스학회지
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    • 제29권4호
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    • pp.221-233
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    • 1999
  • Prospect Hill Virus (PHV) is the well known serotype of hantavirus, a newly established genus in family Bunyaviridae. Extensive studies have upheld the original view of PHV genetics with three genes such as nucleocapsid (N) protein, envelope proteins (G1, G2) and RNA dependent RNA polymerase. In this study, we report the existence of additional gene that is encoded in an overlapping reading frame of the N protein gene within S genome segment of PHV. This gene is expected to encode a nonstructural small (NSs) protein and it seems to be only found in PHV infected cell. The presence and synthesis of NSs protein could be demonstrated in the cell infected with PHV using anti-peptide sera specific to the predicted amino acid sequence deduced from the second open reading frame. Ribosomal synthesis of this protein appears to occur at AUG codon at the 83rd base of S genome segment, downstream of N protein initiation codon. This protein is small in size (10.4 KDa) and highly basic in nature. The expression strategy of NSs protein appears that a signal mRNA is used to translate both N and NSs protein in PHV infected cell. 10 KDa protein in virus infected cell lysates can bind to mimic dsRNA. This fact strongly suggests that NSs protein may be involved in virus replication on late phase of viral life cycle.

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산업폐기물인 비지로부터 식품첨가물로 이용할 수 있는 단백질 가수분해물의 생산 (Production of Protein Hydrolyzate, that can be used as Food Additives, from Okara)

  • 우은열;김민정;신원선;이경애;김강성
    • 한국식품과학회지
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    • 제33권6호
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    • pp.769-773
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    • 2001
  • 두유에서 발생되는 비지와 대두의 일반성분과 단백질의 추출률을 비교하기 위해 시간, 온도, pH별로 분석하였고, 단백질 분자들 사이의 상호작용에 관여하는 물질 urea, SDS, 2-mercaptoethanol를 사용하여 비지단백질의 insolubilization mechamism을 조사하였다. 또한 enzyme modification으로 기능성을 향상시켜 식품소재로서의 이용 가능성에 대해 분석하였다. 비지와 대두는 각각 37.3%, 42.5%의 단백질을 함유하고 있으며 비지는 생산공정 증의 과도한 열처리에 의하여 극히 낮은 용해도를 나타냈다. 비지단백질의 낮은 용해도는 주로 disulfide bonding에 의한 cross linking에 기인하는 것으로 밝혀졌다. 비지단백질과 대두단백질은 pH 3, pH 4에서 가장 낮은 용해도를 보였다. Carbohydrase와 protease를 첨가하여 단백질의 추출츌을 비교한 결과는 비지와 대두는 carbohydrase에서 미세한 반응을 보여 단백질의 용해도에 큰 영향을 주지 못하였으나 여러 protease 가운데 Alcalase는 비지단백질의 용해도를 급격히 증가시켰다.

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Characterization of jute fibre reinforced pine rosin modified soy protein isolate green composites

  • Sakhare, Karishma M.;Borkar, Shashikant P.
    • Advances in materials Research
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    • 제11권3호
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    • pp.191-209
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    • 2022
  • Very slow degradation of synthetic based polymers has created a severe environmental issue that increased awareness towards research in polymers of biodegradable property. Soy protein isolate (SPI) is a natural biopolymer used as matrix in green composites but it has limitations of low mechanical properties and high water sensitivity. To enhance mechanical properties and reduce water sensitivity of Jute-SPI composites, SPI was modified with pine rosin which is also a natural cross-linking agent. 30% glycerol on the weight basis of a matrix was used as a plasticizer. The fibre volume fraction was kept constant at 0.2 whereas the pine rosin in SPI ranged from 5% to 30% of the matrix. The effects of pine rosin on mechanical, thermal, water sensitivity and surface morphology have been characterized using various techniques. The mechanical properties and water absorbency were found to be optimum for 15% pine rosin in Jute-SPI composite. Therefore, Jute-SPI composite without pine rosin and with 15% pine rosin were chosen for investigation through characterization by Fourier transforms infrared spectroscopy (FTIR), Thermo-gravimetric analysis (TGA), X-Ray diffraction (XRD) and Scanning electron microscope (SEM). The surface morphology of the composite was influenced by pine rosin which is shown in the SEM image. TGA measurement showed that the thermal properties improved due to the addition of pine rosin. Antimicrobial test showed antimicrobial property in the composite occurring 15% pine rosin. The research paper concludes that the modification of SPI resin with an optimum percentage of pine rosin enhanced mechanical, thermal as well as water-resistant properties of jute fibre reinforced composites.