• Title/Summary/Keyword: E3KARP

Search Result 4, Processing Time 0.021 seconds

Biochemical and Biophysical Characterizations of the Interaction between Two PDZ Adapter Proteins NHERF and E3KARP in vitro

  • Hwang, Eun-Young;Jeong, Mi-Suk;Jang, Se-Bok
    • Bulletin of the Korean Chemical Society
    • /
    • v.31 no.11
    • /
    • pp.3241-3246
    • /
    • 2010
  • NHERF ($Na^+/H^+$ exchanger regulatory factor) and E3KARP (NHE3 kinase A regulatory protein) play important roles in membrane targeting, trafficking and sorting of ion channels, transmembrane receptors and signaling proteins in many tissues. Each of these proteins contains two PDZ (PSD-95/Dlg-1/ZO-1) domains, which mediate the assembly of transmembrane and cytosolic proteins into functional signal transduction complexes. The interaction between NHERF and E3KARP was investigated by surface plasmon resonance spectroscopy (BIAcore), fluorescence measurement, His-tagged pull-down experiment, and size-exclusion column (SEC) chromatography. BIAcore experiments revealed that NHERF bound to E3KARP with an apparent $K_D$ of 7 nM. Fluorescence emission spectra of the NHERF-E3KARP complex suggested that the tight interaction between these proteins was accompanied by significant conformational changes in one or both. The CD spectra of NHERF and E3KARP show that the conformational changes of these proteins were dependent on pH and temperature. These results implicate that the NHERF-E3KARP complex allows intracellular signaling complexes to form through PDZ-PDZ interactions.

Coculture of Schwann Cells and Neuronal Cells for Myelination in Rat

  • Kim, Ji-Young;Choi, Chang-Shik;Hong, Seong-Karp
    • Rapid Communication in Photoscience
    • /
    • v.3 no.3
    • /
    • pp.48-49
    • /
    • 2014
  • For in vitro myelination system, Schwann cells and neuronal cells of rat were cocultured. Schwann cells and neuronal cells, respectively, were obtained from dorsal root ganglion of rat embryos (E15). This method includes four steps: first step of suspension of the embryonic dorsal root ganglion cells, second step of addition of anti-mitotic cocktail, third step of purification of dorsal root cells, and fourth step of addition of Schwann cells to dorsal root ganglion cells. We made a highly purified population of myelination in a short period through this procedure and identified myelination basic protein using antibody of myelination basic protein.

Coculture of Schwann Cells and Neuronal Cells for Myelination in Rat (랫트에서 수초화를 위한 슈반세포와 뉴런세포의 공동배양)

  • Kweon, Tae-Dong;Sa, Young-Hee;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
    • /
    • 2014.05a
    • /
    • pp.822-825
    • /
    • 2014
  • For in vitro myelination system, Schwann cells and neuronal cells of rat were cocultured. Schwann cells and neuronal cells, respectively, were obtained from dorsal root ganglion of rat embryos (E15). This method includes four steps: first step of suspension of the embryonic dorsal root ganglion cells, second step of addition of anti-mitotic cocktail, third step of purification of dorsal root cells, and fourth step of addition of Schwann cells to dorsal root ganglion cells. We made a highly purified population of myelination in a short period through this procedure and identified myelination basic protein using antibody of myelination basic protein.

  • PDF

AN EXPERIMENTAL STUDY OF THE IRRADIATION EFFECTS ON THE CAPILLARY AND ENDOTHEILIAL CELL OF THE RAT SUBMANDIBULAR GLAND (방사선조사가 악하선 미세혈관과 내피세포에 미치는 영향에 관한 실험적 연구)

  • Yoo Young-Ah;Sohn Jeong-Ick;Choi Mi;Bae Yong-Chul;Choi Karp-Shik
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
    • /
    • v.24 no.1
    • /
    • pp.67-77
    • /
    • 1994
  • The purpose of this study was to investigate the irradiatiion effects on the capillary and endothelial cell in the submandibular gland. Sprague-Dawley strain male rats were singly irradiated to their neck region with the dose of 5Gy by 6MV X-irradiation and sacrificed on the 6 hours, 12 hours, 1, 3, 7, and 14days after irradiation. The authors observed the histological changes of the capillary at H & E and PAS staining under a light microscope, and also observed the ultrastructural changes of the endothelial cell using a transmission electron microscope. The obtaining results were as follows: 1. In the light microscopic examination, the capillary density was slightly increased on the 1day after irradiation, and increased until the 7 days after irradiatiion. After then, capillary density was apparently decreased. 2. The reaction to PAS staining at acinar cells was decreased on the 6 hours after irradiation, and recovered on the 7days after irradiation. But reaction was decreased on the 14days after irradiation agan, after then, gradually recovered with days. 3. In the transmission electron microscopic examination, mild proliferation of cytoplasmic process of the endothelial cell and reduction in luminal size were observed just after irradiation. After then, nuclear degeneration, marked proliferation of cytoplasmic process, thickened basal lamina, and numerous cytoplasmic vesicles were observed on the 1day after irradiation. These changes were recovered to normal on the 14days after 5Gy group, but not with 10Gy irradiation group. And destruction of endothelial cell and loss of basal lamina were not observed in both groups. 4. From the above results, reduction in luminal size, proliferation of cytoplasmic process and thickening of basal lamina were observed as the irradiation effects on the capillary and endothelial cell of the submandibular gland. And also, these changes may induce increase in capillary number and endothelial permeability by means of increase of cytoplasmic vesicle formation. The changes appeared earlier and more prominent in 10Gy irradiated group than in 5Gy irradiated group.

  • PDF