• Title/Summary/Keyword: GPI

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Sorting of the Human Folate Receptor in MDCK Cells

  • Kim, Chong-Ho;Park, Young-Soon;Chung, Koong-Nah;Elwood, P.C.
    • BMB Reports
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    • v.37 no.3
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    • pp.362-369
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    • 2004
  • The human folate receptor (hFR) is a glycosylphosphatidylinositol (GPI) linked plasma membrane protein that mediates delivery of folates into cells. We studied the sorting of the hFR using transfection of the hFR cDNA into MDCK cells. MDCK cells are polarized epithelial cells that preferentially sort GPI-linked proteins to their apical membrane. Unlike other GPI-tailed proteins, we found that in MDCK cells, hFR is functional on both the apical and basolateral surfaces. We verified that the same hFR cDNA that transfected into CHO cells produces the hFR protein that is GPI-linked. We also measured the hFR expression on the plasma membrane of type III paroxysmal nocturnal hemoglobinuria (PNH) human erythrocytes. PNH is a disease that is characterized by the inability of cells to express membrane proteins requiring a GPI anchor. Despite this defect, and different from other GPI-tailed proteins, we found similar levels of hFR in normal and type III PNH human erythrocytes. The results suggest the hypothesis that there may be multiple mechanisms for targeting hFR to the plasma membrane.

Expression of Lymphocyte ADP-ribosyltransferase in Rat Mammary Adenocarcinoma Cells (임파구 ADP-ribosyltransferase의 rat mammary adenocarcinoma cell에서의 발현)

  • 김현주
    • Journal of Life Science
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    • v.8 no.1
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    • pp.102-108
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    • 1998
  • The nascent from of glycosylphosphatidylinositol (GPI)-anchored proteins possesses both amino and carboxy terminal hydrophobic signal sequences to direct processing in the endoplasmic reticulum (ER). Following cleavage of the amino-terminal signal peptide, the carboxy-terminal peptide is processed. Previously, mouse lymphocyte NDA: agrinine ADP-ribosyltransferase (Yac-1) was cloned and the deduced amino acid sequence of the Yac-1 transferase contained hydrophobic amino and carboxy termini, consistent with known signal sequences of GPI-anchored proteins. This tranferase was present on the surface of NMU (rat mammary adenocarcinoma) cells transfected with the wildtype cDNA and was released with phosphatidylinositol-specific phosphilpase C. Expression of the mutant protein, lacking the carboxy terminal hydrophobic sequence, resulted in the peoduction of soluble, secreted from of the transferase. This result shows that carboxy terminal sequence is important for GPI-attachment.

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Spontaneous Release of Glycosylphosphatidylinositol (GPI)-anchored Renal Dipeptidase from Porcine Renal Proximal Tubules

  • Park, Sung-Wook;Kang, Bok-Yun;Yoon, Hyun-Joong;Park, Eun-Mi;Choi, Kyong;Lee, Hwang-Hee Blaise;Hooper, Nigel M.;Park, Haeng-Soon
    • Archives of Pharmacal Research
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    • v.25 no.1
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    • pp.80-85
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    • 2002
  • The incubation of porcine renal proximal tubules (PTs) resulted in the release of the Glycosylphosphatidylinositol (GPI)-anchored renal dipeptidase (RDPase, EC 3. 4. 13. 19) from the membrane after a lag period of approximately 6 hours. This spontaneous release of RDPase from the membrane was inhibited by antibiotics. When the incubation supernatant was added back to fresh PTs, both the antibiotic inhibition of RDPase release and the lag period disappeared. The released RDPase reacted with an anti-cross reacting determinant antibody indicating the presence of the Ins (1, 2-cyc)P. These results suggest that bacteria in the PTs, when incubated, grow find Secrete a phosphatidylinmsitol-specific phospholipase C (PIPLC). This enzyme then hydrolyses the GPI-anchored RDPase and is transferable. RDPase was purified following its release from the membrane by this simple and inexpensive method which may also be applied to other GPI-anchored proteins.

Expression of Folate Receptor Protein in CHO Cell Line

  • Kim, Chong-Ho;Park, Seung-Taeck
    • Biomedical Science Letters
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    • v.14 no.4
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    • pp.203-210
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    • 2008
  • One of cell surface receptor proteins, human folate receptor (hFR) involves in the uptake of folates through cell membrane into cytoplasm, and is anchored to the plasma membrane by a fatty acid linkage, which has been identified in some cells as a glycosylphosphatidylinositol (GPI)-tailed protein with a molecular mass of about 40 kDa. The hFR is released by phosphatidylinositol phospholipase C (PI-PLC) because it contains fatty acids and inositol on the GPI tail. Caveolin decorates the cytoplasmic surface of caveolae and has been proposed to have a structural role in maintaining caveolae. It is unknown whether caveolin is involved in targeting, and is necessary for the function of GPI-tailed proteins. To compare the ability of folic acid binding, internalization and expression of hFR, and the effect of caveolin at the both apical and basolateral side of cell surfaces in Chinese hamster ovary (CHO) clone cells overexpressed the hFR and/or caveolin. Our present results suggest a possibility that the overexpression of caveolin does not be involved in expression of hFR, but plays a role as a factor in PI-PLC releasing kinetics, and for a regulation of formation, processing and function of hFR in CHO clone cells overexpressed cavcolin.

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Deep Brain Stimulation of the Globus Pallidus in a 7-Year-Old Girl with DYT1 Generalized Dystonia

  • Jin, Seon Tak;Lee, Myung Ki;Ghang, Ju Young;Jeon, Seong Man
    • Journal of Korean Neurosurgical Society
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    • v.52 no.3
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    • pp.261-263
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    • 2012
  • The experience of pediatric deep brain stimulation (DBS) of the globus pallidus internus (GPi) in the treatment of early-onset DYT1 generalized dystonia is still limited. Here, we report the surgical experience of bilateral GPi-DBS under general anesthesia by using microelectrode recording in a 7-year-old girl with early-onset DYT1 generalized dystonia. Excellent improvement of her dystonia without neurological complications was achieved. This case report demonstrates that GPi-DBS is an effective and safe method for the treatment of medically refractory early-onset DYT1 generalized dystonia in children.

Water Quality Management System for a Farm Village Stream -watershed monitoring and the system design- (농촌마을 하천의 수질관리 시스템 - 시험유역 조사 및 시스템 설계 -)

  • 정하우;최진용
    • Journal of Korean Society of Rural Planning
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    • v.2 no.2
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    • pp.109-117
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    • 1996
  • The purpose of this study Is to develop water quality management system fort a farm village stream. The framework design of the system and the ecological monitoring of a test watershed were carried out, The system consists of GIS(Geographic Information System ), database, pollution source management, water quality and hydrologic analysis. Suri watershed located on Idong, Yongin city, Kyunggi Province, was selected as the test watershed for the application of the system. The fifteen's monitoring stations were chooses at up- and down-stream of the watershed. The results of an aquatic ecological monitoring were analyzed by the GPI(Group Pollution Index) method. The GPI revealed that water quality was varied within the stream. GPI and DO map for the watershed stream were developed, These maps facilitated to analyze the spatial distribution of the water quality.

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Comparisons of Isozyme Patterns in Pythium Species and Application to Pythium Systematics (Isozyme을 이용한 Pythium species의 비교 및 Pythium systematics에의 이용)

  • Lee, Youn-Su
    • The Korean Journal of Mycology
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    • v.21 no.4
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    • pp.293-300
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    • 1993
  • For the enzymes AAT, GmDH, ME, GPI, LDH and IDH, nine, seven, four, nine, seven, and four different phenotypes, respectively, were observed. All six isolates of an unidentified sterile Pythium sp. isolated from field soil showed the same band positions for all six enzymes compared. These phenotypes were not similar to any of the known Pythium species. Two isolates of unknown Pythium species (145 and 299) showed the same band positions for all six enzymes. The phenotypes for all three unknown Pythium spp. were different from the other species in the experiment. Five isolates of P. heterothallicum showed the same band positions for all enzymes compared except one enzyme, lDH. Two isolates of P. torulosum showed the same band petitions for enzymes AAT, GmDH and ME, and three isolates of P. totulosum showed the same positions for enzymes GPI, LDH, and IDH. Single isolates of P. spinosum and P. irregulare showed the same band positions for enzymes AAT, GmDH and GPI. In conclusion, sterile types of Pythium species showed 100% similarities among themselves but did not show any similarity with all isolates of P. heterothallicum and P. spinosum isolate, and showed very low similarities with other isolates in general except with unknown Pythium isolate 306. Similarity levels between different species were low in general with few exceptions.

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Continuous Sliding Mode Control for Permanent Magnet Synchronous Motor Speed Regulation Systems Under Time-Varying Disturbances

  • Wang, Huiming;Li, Shihua;Yang, Jun;Zhou, XingPeng
    • Journal of Power Electronics
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    • v.16 no.4
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    • pp.1324-1335
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    • 2016
  • This article explores the speed regulation problem of permanent magnet synchronous motor (PMSM) systems subjected to unknown time-varying disturbances. A continuous sliding mode control (CSMC) technique is introduced for the speed loop to enhance the robustness of PMSM systems and eliminate the chattering phenomenon caused by high-frequency switch function in the conventional control law. However, the high control gain of the CSMC law in the presence of strong disturbances leads to large steady-state speed fluctuations for PMSM systems. In many application fields, PMSM systems are affected by time-varying disturbances instead of constant disturbances. For example, electric bicycles are usually affected by changing environmental disturbances, including wind speeds, road conditions, etc. These disturbances may be in the form of constant, ramp, and parabolic disturbances. Hence, a generalized proportional integral (GPI) observer is employed to estimate these types of disturbances. Then, the disturbance estimation method and the aforementioned CSMC method are combined to establish a composite sliding mode control method called the CSMC+GPI method for the speed loop of PMSM systems. Contrary to the conventional sliding mode control technique, the proposed method completely eliminates the chattering phenomenon caused by the switching function in the conventional control law. Moreover, a small control gain for the CSMC+GPI method is chosen by feed-forwarding estimated values to the speed controller. Hence, the steady-state speed fluctuations are small. The effectiveness of the proposed control scheme is verified by simulation and experimental result.

Sorting and Function of the Human Folate Receptor Is Independent of the Caveolin Expression in Fisher Rat Thyroid Epithelial Cells

  • Kim, Chong-Ho;Park, Young-Soon;Chung, Koong-Nah;Elwood, Patrick C.
    • BMB Reports
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    • v.35 no.4
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    • pp.395-402
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    • 2002
  • Caveolae are small, flask-shaped, non-clathrin coated invaginations of the plasma membrane of many mammalian cells. Caveolae have a coat that includes caveolin. They have been implicated in numerous cellular processes, including potocytosis. Since the human folate receptor (hFR) and other glycosyl-phosphatidylinositol (GPI)-tailed proteins have been co-localized to caveolae, we studied the caveolin role in the hFR function by transfecting hFR and/or caveolin cDNA into Fischer rat thyroid epithelial (FRT) cells that normally do not express detectable levels of either protein. We isolated and characterized stable clones as follows: they express (1) high levels of caveolin alone, (2) hFR and caveolin, or (3) hFR alone. We discovered that hFR is correctly processed, sorted, and anchored by a GPI tail to the plasma membrane in FRT cells. No difference in the total folic acid binding or cell surface folic acid binding activity were found between the FRT cells that were transfected with hFR, or cells that were transfected with hFR and caveolin. The hFR that was expressed on the cell surface of clones that were transfected with hFR was also sensitive to phosphatidylinositol-specific phospholipase C (PI-PLC) release, and incorporated radiolabeled ethanolamine that supports the attachment of a GPI-tail on hFR. We conclude that the processing, sorting, and function of hFR is independent on the caveolin expression in FRT cells.

Characterization of Phosphatidylinositol Glycan, Class K (PIGK) Gene and Analysis of Association with Quantitative Traits in Pigs (돼지 Phosphatidylinositol Glycan, Class K (PIGK) 유전자의 동정과 양적형질과의 연관성 분석)

  • Lim, H.T.;Kim, J.H.;Choi, B.H.;Lee, S.H.;Park, E.W.;Kim, T.H.;Cho, I.C.;Oh, S.J.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.47 no.2
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    • pp.167-176
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    • 2005
  • PIGK(phosphatidylinositol glycan, class K) is a subunit of GPI transamidase that cleaves the signal peptide in proproteins and replaces it with GPI. In addition, the structure and synthesis of GPI are critically involved in some of the cellular actions of insulin. Therefore, PIGK would be essential for mammalian development and many specific cellular functions as well as for metabolic activity of insulin associated with GPI. Two types of" full-length cDNAs of porcine PIGK were cloned through RT-PCR and RACE experiments. One is thought to be a normal form(consist of 395 amino acids) and the other is considered as an alternative spliced form(consist of 371 amino acids) which contains additional 63 bps in intron 7. Since a stop codon was contained within the insertion, the spliced form has a shorter coding sequence than that of normal form. A missense mutation (T314I) in exon 6 was detected and used for genotyping to estimate association with the growth and fat deposition traits for 545 $F_2$ animals(Korean native boars ${\times}$ Landrace). From the PCR-RFLP analysis using HpyCH4III, CT genotype showed highly significant relationship(P< 0.01) with carcass fat contents.