• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.458-462
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• 2001

The localization of paclitaxel was investigated in suspension culture cells of Taxus chinensis. Over 93% of the cell-associated paclitaxel were detected throughout the entire culture period. Intracellular localization of paclitaxel over the culture time was analyzed further by cell fractionation for days 21 and 42. Paclitaxel contents in intracellular organelles were decreased at day 42, while the content in the cell wall fraction was increased at day 42 compared to the value for day 21. The localization of paclitaxel in the cell wall was confirmed by using the immunocytochemical method with the aid of a confocal laser scanning microscope.

• Journal of Plant Biology
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• 제28권3호
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• pp.225-232
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• 1985

Intracellular localization and the developmental changes in activities of uricase and allantoinase during germination were investigated with the cotyledons of rape(Brassica napus L.) seedlings. The development anddisappearance of uricase activity took place independently of light, but allantoinase activity was increased by light. The temporal pattern of uricase activity showed that uricolysis was actively taking place in the cotyledons during their early stages of germination. While uricase can be localized in the microbody fraction isolated from crude organelle extracts of the cotyledons by density gradient centrifugation, most of the allantoinase activity found in the microbody fraction did not appear to be an integral part of the microbody.

• Bulletin of the Korean Chemical Society
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• 제30권9호
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• pp.1981-1984
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• 2009

Subcellular localization of protein kinase often plays an important role in determining its activity and specificity. Protein kinase C (PKC), a family of multi-gene protein kinases has long been known to be translocated to the particular cellular compartments in response to DAG or its analog phorbol esters. We used C-terminal green fluorescent protein (GFP) fusion proteins of PKC isoforms to visualize the subcellular distribution of individual PKC isoforms. Intracellular localization of PKC-GFP proteins was monitored by fluorescence microscopy after transient transfection of PKC-GFP expression vectors in the HeLa cells. In unstimulated HeLa cells, all PKC isoforms were found to be distributed throughout the cytoplasm with a few exceptions. PKC$\theta$ was mostly localized to the Golgi, and PKC$\gamma$, PKC$\delta$ and PKC$\eta$ showed cytoplasmic distribution with Golgi localization. DAG analog TPA induced translocation of PKC-GFP to the plasma membrane. PKC$\alpha$, PKC$\eta$ and PKC$\theta$ were also localized to the Golgi in response to TPA. Only PKC$\delta$ was found to be associated with the nuclear membrane after transient TPA treatment. These results suggest that specific PKC isoforms are translocated to different intracellular sites and exhibit distinct biological effects.

• Fisheries and Aquatic Sciences
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• 제6권3호
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• pp.135-139
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• 2003

During routine survey of Pacific oyster (Crassostrea gigas) collected from Tongyoung area in southern coast of Korea, histological examination revealed that a intracellular microorganisms infected the digestive gland of the oyster. They infected hepatopancreatic cells extensively. The size of intracellular microorganism was of 45 to 86nm in diameter and 200nm to more thar 500nm in length. They were pleomorphic. The morphological characteristic of intracellular microorganisms lacked cell wall and was bounded by the plasma membrane. They contained typical prokaryotic ribosomes and fibrillar DNA-like strands. No additional internal structure has been observed. Based on the lack of cell wall and the cellular localization, the intracellular microorganism is considered as a Mycoplasma-like organism.

• BMB Reports
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• 제43권10호
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• pp.656-663
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• 2010

Amyloid precursor protein (APP), which is critically involved in the pathogenesis of Alzheimer's disease (AD), is cleaved by gamma/epsilon-secretase activity and results in the generation of different lengths of the APP Intracellular C-terminal Domain (AICD). In spite of its small size and short half-life, AICD has become the focus of studies on AD pathogenesis. Recently, it was demonstrated that AICD binds to different intracellular binding partners ('adaptor protein'), which regulate its stability and cellular localization. In terms of choice of adaptor protein, phosphorylation seems to play an important role. AICD and its various adaptor proteins are thought to take part in various cellular events, including regulation of gene transcription, apoptosis, calcium signaling, growth factor, and $NF-{\kappa}B$ pathway activation, as well as the production, trafficking, and processing of APP, and the modulation of cytoskeletal dynamics. This review discusses the possible roles of AICD in the pathogenesis of neurodegenerative diseases including AD.

• 식물조직배양학회지
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• 제27권5호
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• pp.407-409
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• 2000

Outer envelope membrane proteins of chloroplasts encoded by the nuclear genome are transported without the N-terminal transit peptide. Here, we investigated the targeting mechanism of AtOEP7, an Arabidopsis homolog of small outer envelope membrane proteins in vivo. AtOEP7 was expressed transiently in protoplasts or stably in transgenic plants as fusion proteins with GFP. In both cases AtOEP7:GFP was targeted to the outer envelope membrane when assayed under a fluorescent microscope or by Western blot analysis. Except the transmembrane domain, deletions of the N- or C-terminal regions of AtOEP7 did not affect targeting although a region closed to the C-terminal side of the transmembrane domain affected the targeting efficiency. Targeting experiments with various hybrid transmembrane mutants revealed that the amino acid sequence of the transmembrane domain determines the targeting specificity The targeting mechanism was further studied using a fusion protein, AtOEP7：NLS：GFP, that had a nuclear localization signal. AtOEP7：NLS：GFP was efficiently targeted to the chloroplast envelope despite the presence of the nuclear localization signal. Taken together, these results suggest that the transmembrane domain of AtOEP7 functions as the sole determinant of targeting specificity and that AtOEP7 may be associated with a cytosolic component during translocation to the chloroplast envelope membrane.

• 분석과학
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• 제28권1호
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• pp.65-71
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• 2015

진핵세포내 막성세포소기관들은 각각 고유한 세포의 중요한 기능들을 담당하고 있다. 이들 기관에 분포하는 단백질들은 세포질에서 발현된 후, 정교한 조절에 의해서 다양한 세포내 소기관으로 운송된다. 따라서, 세포내에 존재하는 막성세포소기관의 마커를 개발하고, 이들의 타기팅 기전을 알아내는 것은 세포 생리 및 병리학적 기전 연구에 중요한 도구가 될 수 있다. 본 연구에서는 기존에 보고된 당지질-결합 펩타이드들과 이들의 변형을 통한 세포내 타겟팅을 분석하였다. 그 결과 이러한 당지질-결합 펩타이드들은 미토콘드리아, 원형질막, 골지체로 위치하는 것을 확인할 수 있었으며, 이러한 펩타이드가 세포내 기관을 마커로 이용될 가능성을 확인할 수 있었다. 또한, 원형질막에 타기팅하는 펩타이드 마커의 경우는, 정전기적인 상호작용에 의해 원형질막에 선택적으로 타기팅됨을 알 수 있었다. 본 연구결과를 통해 당지질-결합 펩타이드들이 다양한 세포내 운송과 관련한 연구에 세포소기관의 위치 및 모양을 분석 할 수 있는 마커로 이용될 수 있음을 알 수 있었다.

• 동의생리병리학회지
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• 제31권2호
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• pp.105-110
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• 2017

Prolonged exposure to solar ultraviolet A (UVA) radiation has been known to cause premature skin aging (photo-aging). UVA radiation generates ROS thereby induce degenerative changes of skin such as degradation of dermal collagen, elastic fibers. Matrix metalloproteinases (MMPs), the proteolytic enzymes have been implicated as a major player in the development of UVA-induced photo-aging. Many studies have been conducted to block the harmful effects of UV radiation on the skin. Recently, we are interested in the availability of fermented red ginseng (FRG) as natural matrix metalloproteinases inhibitors (MMPIs). The efficacy difference between red ginseng and FRG has been compared. Both RG and FRG have no cytotoxic effects below the concentration of $300{\mu}g/ml$. Human dermal fibroblasts (HDFs) were pretreated with FRG or RG for 24h, followed by irradiation of UVA. Then, we measured the intracellular ROS production and the expression of MMP, $IL-1{\beta}$ at the mRNA level. We also examined the intracellular localization of $NF-{\kappa}B$ and MMP-9 on the FRG or RG treated and UVA-irradiated HDFs. FRG decreased the intracellular ROS production elicited by UVA. In addition, FRG decreased the mRNA expression of MMP-3, MMP-9, and $IL-1{\beta}$ more efficiently than RG. Furthermore, FRG suppressed the nuclear localization of $NF-{\kappa}B$, and the expression of MMP-9. Taken together, our results suggest that FRG is promising agents to prevent UVA-induced photo-aging by suppressing MMP expression and inflammation.

• Molecules and Cells
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• 제45권11호
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• pp.855-867
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• 2022

For proper function of proteins, their subcellular localization needs to be monitored and regulated in response to the changes in cellular demands. In this regard, dysregulation in the nucleocytoplasmic transport (NCT) of proteins is closely associated with the pathogenesis of various neurodegenerative diseases. However, it remains unclear whether there exists an intrinsic regulatory pathway(s) that controls NCT of proteins either in a commonly shared manner or in a target-selectively different manner. To dissect between these possibilities, in the current study, we investigated the molecular mechanism regulating NCT of truncated ataxin-3 (ATXN3) proteins of which genetic mutation leads to a type of polyglutamine (polyQ) diseases, in comparison with that of TDP-43. In Drosophila dendritic arborization (da) neurons, we observed dynamic changes in the subcellular localization of truncated ATXN3 proteins between the nucleus and the cytosol during development. Moreover, ectopic neuronal toxicity was induced by truncated ATXN3 proteins upon their nuclear accumulation. Consistent with a previous study showing intracellular calcium-dependent NCT of TDP-43, NCT of ATXN3 was also regulated by intracellular calcium level and involves Importin α3 (Imp α3). Interestingly, NCT of ATXN3, but not TDP-43, was primarily mediated by CBP. We further showed that acetyltransferase activity of CBP is important for NCT of ATXN3, which may acetylate Imp α3 to regulate NCT of ATXN3. These findings demonstrate that CBP-dependent acetylation of Imp α3 is crucial for intracellular calcium-dependent NCT of ATXN3 proteins, different from that of TDP-43, in Drosophila neurons.

• Parasites, Hosts and Diseases
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• 제42권4호
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• pp.185-193
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• 2004

Toxoplasma gondii is an obligate intracellular protozoan parasite, which invades a wide range of hosts including humans. The exact mechanisms involved in its invasion are not fully understood. This study focused on the roles of $Ca^{2+}$ in host cell invasion and in T. gondii replication. We examined the invasion and replication of T. gondii pretreated with several calcium modulators, the conoid extrusion of tachyzoites. Calmodulin localization in T. gondii were observed using the immunogold method, and $Ca^{2+}$ levels in tachyzoites by confocal microscopy. In light microscopic observation, tachyzoites co-treated with A23187 and EGTA showed that host cell invasion and intracellular replication were decreased. The invasion of tachyzoites was slightly inhibited by the $Ca^{2+}$ channel blockers, bepridil and verapamil, and by the calmodulin antagonist, calmidazolium. We observed that calcium saline containing A23187 induced the extrusion of tachyzoite conoid. By immunoelectron microscopy, gold particles bound to anti-calmodulin or anti-actin mAb, were found to be localized on the anterior portion of tachyzoites. Remarkably reduced intracellular $Ca^{2+}$ was observed in tachyzoites treated with BAPTA/AM by confocal microscopy. These results suggest that host cell invasion and the intracellular replication of T. gondii tachyzoites are inhibited by the calcium ionophore, A23187, and by the extracellular calcium chelator, EGTA.