• 대한의생명과학회지
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• 제12권3호
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• pp.185-190
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• 2006

Lysyl oxidase (LOX) catalyzes the lysine-derived cross-links of fibrillar collagens and elastin in the extracellular matrix. Recent molecular cloning has revealed existence of a LOX family consisting of LOX and four lysyl oxidase-like proteins (LOXL, LOXL2, LOXL3 and LOXL4). Pathological conditions associated with impaired LOX activity in several heritable and acquired disorders lead to severe structural and functional abnormalities of cardiovascular tissues, such as occlusion of coronary arteries and aneurysms, suggesting an essential role for the LOX family proteins in the maintenance of the cardiovascular system. However, the specific roles of the lysyl oxidase-like proteins in normal and pathological conditions of the cardiovascular tissues have not been established yet. Here, I report that LOXL3, a novel member of the LOX family, is predominantly expressed in the aorta, with an amine oxidase activity toward collagen and elastin, suggesting an essential role of LOXL3 in the development and maintenance of the aorta.

• BMB Reports
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• 제44권1호
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• pp.22-27
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• 2011

A series of elastin-like proteins, $SKGPG[V(VKG)_3VKVPG]_n$-(ELP1-90)WP (n = 1, 2, 3, and 4), were biosynthesized based on the hydrophobic and lysine linkage domains of tropoelastin. The formation of self-assembled hydrophobic aggregates was monitored in order to determine the influence of cationic segments on phase transition properties as well as the sensitivity to changes in salt and pH. The thermal transition profiles of the proteins fused with only one or two cationic blocks (n = 1 or 2) were similar to that of the counterpart ELP1-90. In contrast, diblock proteins that contain 3 and 4 cationic blocks displayed a triphasic profile and no transition, respectively. Upon increasing the salt concentration and pH, a stimulus-induced phase transition from a soluble conformation to an insoluble aggregate was observed. The effects of cationic segments on the stimuli sensitivity of cationic bimodal ELPs were interpreted in terms of their structural and molecular characteristics.

• Bulletin of the Korean Chemical Society
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• 제15권3호
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• pp.249-256
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• 1994

Fluorescence time decay of human aorta has been measured at 380, 440, 480 nm using 320 nm excitation and time-correlated single photon counting technique. Fluorescence decay was found to be nonexponential at all emission frequencies. The normal and diseased sample showed significantly different fluorescence behaviors at 380 nm while this time decay difference was decreased in the fluorescence at 440 and 480 nms. The decay data were multiexponential and were analyzed with two exponential decay constants. The fluorescence decays were compared with and analyzed in terms of collagen and elastin.

• 대한화장품학회지
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• 제34권3호
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• pp.217-223
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• 2008

피부에서 엘라스틴(elastin)은 중요한 탄력섬유의 구성성분 중의 하나이다. 피부주름(skin wrinkle) 형성은 콜라겐(collagen)의 합성과 분해가 중요한 요인으로 작용한다고 알려져 있지만, 최근 많은 연구에서 엘라스틴의 재형성과 분해 또한 주름형성 기전에서 중요한 작용을 하는 것으로 보고되고 있다. 엘라스타제(elastase)는 엘라스틴을 분해하는 일종의 메탈로프로테이나제(metalloproteinase)이며, 자외선 B (ultraviolet B, UVB) 조사 후에 활성이 증가하는 것으로 알려져 있다. 따라서 증가된 엘라스타제의 활성은 피부의 탄력성 감소와 주름 형성의 주요한 원인이 될 것이다. 본 연구에서는 항산화 효과가 있으며, 각종 견과류나 과일에 함유된 폴리페놀(polyphenol)성분인 탄닌산(tannic acid)을 사람의 섬유아세포(CCD-25Sk fibroblasts)에 처리하여 엘라스타제 활성과 트로포엘라스틴 생성에 미치는 영향을 조사하였다. 탄닌산은 농도의존적으로 사람의 섬유아세포 엘라스타제 활성을 유의성 있게 억제시켰다. 그러나 트로포엘라스틴 합성이나 발현증가에는 유의성 있는 효과를 보이지 않았다. 이러한 결과들로부터 탄닌산은 엘라스타제의 활성을 억제시켜 엘라스틴의 3차원적 구조를 유지하는데 기여하는 것으로 사료된다. 따라서 탄닌산은 주름생성을 억제하는 물질로 개발 가능할 것으로 기대된다.

• Applied Microscopy
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• 제24권2호
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• pp.37-47
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• 1994

Using mouse tissue, we studied electron opacity effect of tannic acid for transmission electron microscopic staining. Tannic acid-glutaraldehyde in 0.1M phosphate buffer was used as a fixative. To compare with this we have tested another method consisting of heavy metal staining after treatment of tannic acid in sodium tetraborate (borax) on glutaraldehyde-fixed sections. We have achieved equally consistent electron opacity in both methods. The elastin, collagen, basal lamina of skin and gap junctions of the epithelial cells gave excellent results, while it was good for glycogen, cilia, and plasma. Also fat cells and lipid droplets gave good preservation when tannic acid was added in the fixative. However, prolonged fixation in tannic acid-added fixative was hazardous for further processing, i.e., sectioning problem and deep electron opacity background.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1998년도 한국생물과학협회 학술발표대회
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• pp.313.1-313
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• 1998

No Abstract, See Full Text

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVII)
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• pp.562-562
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• 2005

• Journal of Microbiology and Biotechnology
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• 제17권11호
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• pp.1751-1757
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• 2007

Elastin-like polypeptides (ELPs) undergo a reversible inverse phase transition upon a change in temperature. This thermally triggered phase transition allows for a simple and rapid means of purifying a fusion protein. Recovery of ELPs-tagged fusion protein was easily achieved by aggregation, triggered either by raising temperature or by adding salt. In this study, levansucrase has been used as a model enzyme in the development of a simple one-step purification method using ELPs. The levansucrase gene cloned from Pseudomonas aurantiaca S-4380 was tagged with various sizes of ELPs to functionally express and optimize the purification of levansucrase. One of two ELPs, ELP[V-20] or ELP[V-40], was fused at the C-terminus of the levansucrase gene. A levansucrase-ELP fusion protein was expressed in Escherichia coli $DH5{\alpha}$ at $37^{\circ}C$ for 18 h. The molecular masses of levansucrase-ELP[V-20] and levansucrase-ELP[V-40] were determined as 56 kDa and 65 kDa, respectively. The phase transition of levansucrase-ELP[V-20] occurred at $20^{\circ}C$ in 50 mM Tris-Cl (pH 8) buffer with 3 M NaCl added, whereas the phase transition temperature ($T_t$) of levansucrase-ELP[V-40] was $17^{\circ}C$ with 2 M NaCl. Levansucrase was successfully purified using the phase transition characteristics of ELPs, with a recovery yield of higher than 80%, as verified by SDS-PAGE. The specific activity was measured spectrophotometrically to be 173 U/mg and 171 U/mg for levansucrase-ELP[V-20] and levansucrase-ELP[V-40], respectively, implying that the ELP-tagging system provides an efficient one-step separation method for protein purification.

• 대한의생명과학회지
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• 제19권1호
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• pp.41-47
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• 2013

Pluripotent stem cells (PSCs) have lots of potential in biomedical sciences owing to its potential to differentiate into any kind of cells in the body. However, it is still a challenge to culture PSCs on a large scale for application to regenerative medicine. Herein, we introduce a synthetic polymer that enables large-scale suspension culture of human PSCs. By employing suspension culture, it became unnecessary to use conventional substrata such as mouse embryonic fibroblast (MEF) or Matrigel$^{TM}$, which are believed to be main causative sources of xenogeneic contamination in cultured human PSCs in vitro. Human PSCs were cultured in the medium in which elastin-like polypeptide (ELP) dissolved. The ELP in the medium became harden as temperature increases by transforming the medium into a semi-solid gel that supported growth of human PSCs in suspension. Gel-sol transition temperature of ELP can be adjusted by modifying the peptide sequence in which 5 amino acids, Val-Pro-Gly-Xaa-Gly, repeated sequentially. We constructed 3D suspension media having transition temperature around $33{\sim}35^{\circ}C$ using an ELP consisted of 40, 60, or 80 repeats of a monomer, which was Val-Pro-Gly-Val-Gly. Among the ELPs, ELP80 was chosen as the best ELP to support growth of human PSCs in suspension culture. This result suggests that the ELP80 can be a medium component for culturing human PSCs in large-scale.

• Molecules and Cells
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• 제41권3호
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• pp.198-206
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• 2018

Aortic dissection (AD) is a catastrophic disease with high mortality and morbidity, characterized with fragmentation of elastin and loss of smooth muscle cells. Although AD has been largely attributable to polymorphisms defect in the elastin-coding gene, tropoelastin (TE), other undermined factors also appear to play roles in AD onset. Here, we investigated the effects of post-transcriptional control of TE by microRNAs (miRNAs) on elastin levels in aortic smooth muscle cells (ASMC). We found that miR-144-3p is a miRNA that targets TE mRNA in both human and mouse. Bioinformatics analyses and dual luciferase reporter assay showed that miR-144-3p inhibited protein translation of TE, through binding to the 3'-UTR of the TE mRNA. Interestingly, higher miR-144-3p levels and lower TE were detected in the ASMC obtained from AD patients, compared to those from non-AD controls. In a mouse model for human AD, infusion of adeno-associated viruses (serotype 6) carrying antisense for miR-144-3p (asmiR-144-3p) under CAG promoter significantly reduced the incidence and severity of AD, seemingly through enhancement of TE levels in ASMC. Thus, our data suggest an essential role of miR-144-3p on the pathogenesis of AD.