• Title/Summary/Keyword: membrane chemistry

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Subcellular Localization of Diacylglycerol-responsive Protein Kinase C Isoforms in HeLa Cells

  • Kazi, Julhash U.;Kim, Cho-Rong;Soh, Jae-Won
    • Bulletin of the Korean Chemical Society
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    • v.30 no.9
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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.

Effect of Ganglioside $G_{M3}$ on the Erythrocyte Glucose Transporter (GLUT1): Conformational Changes Measured by Steady-State and Time-Resolved Fluorescence Spectroscopy

  • Yoon, Hae-Jung;Lee, Min-Yung;Jhon, GiI-Ja
    • BMB Reports
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    • v.30 no.4
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    • pp.240-245
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    • 1997
  • Interactions between ganglioside $G_{M3}$ and glucose transporter, GLUT1 were studied by measuring the effect of $G_{M3}$ on steady-state and time-resolved fluorescence of purified GLUT1 in synthetic lipids and on the 3-O-methylglucose uptake by human erythrocytes. The intrinsic tryptophan fluorescence showed a GLUT 1 emission maximum of 335 nm, and increased in the presence of $G_{M3}$ by 12% without shifting the emission maximum, The fluorescence lifetimes of intrinsic tryptophan on GLUT1 consisted of a long component of 7.8 ns and a short component of 2,3 ns and $G_{M3}$ increased both lifetime components. Lifetime components were quenched by acrylamide and KI. Acrylarnide-mduced quenching of long-lifetime components was partly recovered by $G_{M3}$ However. KI-induccd quenching of short- and long-lifetime components was not rescued by $G_{M3}$. The anisotropy of 1.6-diphenyl-1.3.5-hexatriene (DPH)-probed dimyristoylphosphatidylcholine (DMPC) model membrane was also increased with $G_{M3}$ incorporation, The transport rate of 3-O-methylglucose increased by 20% with $G_{M3}$ incorporation on the erythrocytes, Therefore, $G_{M3}$ altered the environment of lipid membrane and induced the conformational change of GLUT1.

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Novel Copper(Ⅱ)-Selective Senor Based on a New Hexadentates Schiff's Base

  • Ganjali, Mohammad Reza;Emami, Mehdi;Salavati Niasari, Masoud
    • Bulletin of the Korean Chemical Society
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    • v.23 no.10
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    • pp.1394-1398
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    • 2002
  • A novel copper(II) membrane electrode based on diphenylisocyanate bis(acetylacetone) ethylenediimine (DIBAE), as a new hexadentates Schiff's base was prepared. The electrode exhibited a Nernstian response for Cu$^{2+}$ ions over a wide concentration range (1.0 ${\times}$ 10$^{-1}$ to 1.0 ${\times}$ l0$^{-6}$ M) with a limit of detection of 6.0 ${\times}$ 10$^{-7}$ M (39 ppb). The sensor shows a fast response time (15s) and the membrane can be used for more than 4 months without observing any major deviation. The electrode revealed very good selectivity with respect to many cations including alkali, alkaline earth, transition and heavy metal ions. The proposed sensor could be used in a pH range of 3.0-7.5. It was applied to the direct potentiometric determination of copper in black tea, and in wastewater of copper electroplating samples. The electrode was also used in potentiometric titration of the copper(II) ion with EDTA.

Structure of CT26 in the C-terminal of Amyloid Precursor Protein Studied by NMR Spectroscopy

  • Kang, Dong-Il;Baek, Dong-Ha;Shin, Song-Yub;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.26 no.8
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    • pp.1225-1228
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    • 2005
  • C-terminal fragments of APP (APP-CTs), that contain A$\beta$ sequence, are found in neurotic plaques, neurofibrillary tangles and the cytosol of lymphoblastoid cells obtained from AD patients. CT26, Thr639-Asp664 (TVIVITLVMLKKKQYTSIHH GVVEVD) includes not only the transmembrane domain but also the cytoplasmic domain of APP. This sequence is produced from cleavage of APP by caspase and $\gamma$-secretase. In this study, the solution structure of CT26 was investigated using NMR spectroscopy and circular dichroism (CD) spectropolarimeter in various membrane-mimicking environments. According to CD spectra and the tertiary structure of CT26 determined in TFE-containing aqueous solution, CT26 has an α-helical structure from $Val^{2}\;to\;Lys^{11}$ in TFE-containing aqueous solution. However, according to CD data, CT26 adopts a $\beta$-sheet structure in the SDS micelles and DPC micelles. This result implies that CT26 may have a conformational transition between $\alpha$-helix and $\beta$-sheet structure. This study may provide an insight into the conformational basis of the pathological activity of the C-terminal fragments of APP in the model membrane.

Sulfonated Dextran/Poly(vinyl alcohol) Polymer Electrolyte Membranes for Direct Methanol Fuel Cells

  • Won, Jong-Ok;Ahn, Su-Mi;Cho, Hyun-Dong;Ryu, Ji-Young;Ha, Heung-Yong;Kang, Yong-Soo
    • Macromolecular Research
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    • v.15 no.5
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    • pp.459-464
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    • 2007
  • Polymer electrolyte membranes, featuring ionic channels, were prepared from sulfonated dextran/ poly(vinyl alcohol) (sD/PVA) membranes. A stiff sulfated dextran was chosen as the route for ionic transport, since ionic sites are located along the stiff dextran main chain. The sD/PVA blend membranes were annealed and then chemically crosslinked. The characteristics of the crosslinked sD/PVA membranes were investigated to determine their suitability as proton exchange membranes. The proton conductivity was found to increase with increasing amounts of sD inside the membrane, which reached a maximum and then decreased when the sD content exceeded 30 wt%, while the methanol permeability increased with increasing sD content. The good dispersion of sD inside the membrane, which serves as an ionic channels mimic, played a significant role in proton transportation.