• Title/Summary/Keyword: K-ATPase protein

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Increased 26S proteasome non-ATPase regulatory subunit 1 in the aqueous humor of patients with age-related macular degeneration

  • Lee, Hyungwoo;Choi, Ae Jin;Kang, Gum-Yong;Park, Hyung Soon;Kim, Hyung Chan;Lim, Hyunjung Jade;Chung, Hyewon
    • BMB Reports
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    • v.47 no.5
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    • pp.292-297
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    • 2014
  • Age-related macular degeneration (AMD) is the leading cause of blindness in the world. Evidence indicates that the suppression of the ubiquitin-proteasome system (UPS) contributes to the accumulation of toxic proteins and inflammation in retinal pigment epithelium (RPE), the functional abnormalities and/or the degeneration of which are believed to be the initiators and major pathologies of AMD. To identify new protein associations with the altered UPS in AMD, we used LC-ESI-MS/MS to perform a proteomic analysis of the aqueous humor (AH) of AMD patients and matched control subjects. Six UPS-related proteins were present in the AH of the patients and control subjects. Four of the proteins, including 26S proteasome non-ATPase regulatory subunit 1 (Rpn2), were increased in patients, according to semi-quantitative proteomic profiling. An LC-MRM assay revealed a significant increase of Rpn2 in 15 AMD patients compared to the control subjects, suggesting that this protein could be a biomarker for AMD.

Changes in Physicochemical and Rheological Properties of Fish Meat Paste during Repeated Freezing and Thawing Process (냉동고기풀의 동결-해동 반복에 의한 물리$\cdot$화학적 성질 및 물성변화)

  • CHOI Hyeon-Mee;KIM Mu-Nam;CHUN Soon-Sil;LEE Keun-Tai
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.22 no.3
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    • pp.115-120
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    • 1989
  • Physicochemical changes were investigated during repeated freezing and thawing processes using fish meat paste of alaska pollack (Theragra chalcogramma). During repeated thawing process, the solubility of myofibrillar protein, $Ca^{++}-ATPase$ activity, water holding capacity (WHC), electrophoretic patterns and rheological properties were evaluated at various thawing temperatures. Solubility of myofibrillar protein and $Ca^{++}-ATPase$ activity were decreased with increasing thawing temperatures. Thawing temperatures and the frequency of freezing and thawing processes did not affect WHC significantly. Upon repeated freezing and thawing cycles, electrophoretic patterns showed that only the amount of myosin heavy chain was decreased, whereas the amount of actin remained constant. Young's modulus for viscoelasticity of fish meat pastes increased with increasing thawing temperatures and the value showed maximum at third cycle and decreased thereafter.

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Na/K-ATPase beta1-subunit associates with neuronal growth regulator 1 (NEGR1) to participate in intercellular interactions

  • Cheon, Yeongmi;Yoo, Ara;Seo, Hyunseok;Yun, Seo-Young;Lee, Hyeonhee;Lim, Heeji;Kim, Youngho;Che, Lihua;Lee, Soojin
    • BMB Reports
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    • v.54 no.3
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    • pp.164-169
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    • 2021
  • Neuronal growth regulator 1 (NEGR1) is a GPI-anchored membrane protein that is involved in neural cell adhesion and communication. Multiple genome wide association studies have found that NEGR1 is a generic risk factor for multiple human diseases, including obesity, autism, and depression. Recently, we reported that Negr1-/- mice showed a highly increased fat mass and affective behavior. In the present study, we identified Na/K-ATPase, beta1-subunit (ATP1B1) as an NEGR1 binding partner by yeast two-hybrid screening. NEGR1 and ATP1B1 were found to form a relatively stable complex in cells, at least partially co-localizing in membrane lipid rafts. We found that NEGR1 binds with ATP1B1 at its C-terminus, away from the binding site for the alpha subunit, and may contribute to intercellular interactions. Collectively, we report ATP1B1 as a novel NEGR1-interacting protein, which may help deciphering molecular networks underlying NEGR1-associated human diseases.

CHIP promotes the degradation of mutant SOD1 by reducing its interaction with VCP and S6/S6' subunits of 26S proteasome

  • Choi, Jin-Sun;Lee, Do-Hee
    • Animal cells and systems
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    • v.14 no.1
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    • pp.1-10
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    • 2010
  • Previously we showed that CHIP, a co-chaperone of Hsp70 and E3 ubiquitin ligase, can promote the degradation of mutant SOD1 linked to familial amyotrophic lateral sclerosis (fALS) via a mechanism not involving SOD1 ubiquitylation. Here we present evidence that CHIP functions in the interaction of mutant SOD1 with 26S proteasomes. Bag-1, a coupling factor between molecular chaperones and the proteasomes, formed a complex with SOD1 in an hsp70-dependent manner but had no direct effect on the degradation of mutant SOD1. Instead, Bag-1 stimulated interaction between CHIP and the proteasome-associated protein VCP (p97), which do not associate normally. Over-expressed CHIP interfered with the association between mutant SOD1 and VCP. Conversely, the binding of CHIP to mutant SOD1 was inhibited by VCP, implying that the chaperone complex and proteolytic machinery are competing for the common substrates. Finally we observed that mutant SOD1 strongly associated with the 19S complex of proteasomes and CHIP over-expression specifically reduced the interaction between S6/S6' ATPase subunits and mutant SOD1. These results suggest that CHIP, together with ubiquitin-binding proteins such as Bag-1 and VCP, promotes the degradation of mutant SOD1 by facilitating its translocation from ATPase subunits of 19S complex to the 20S core particle.

Expression, Purification and Characterization of the BLM binding region of human Fanconi Anemia Group J Protein

  • Yeom, Kyuho;Park, Chin-Ju
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.1
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    • pp.22-26
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    • 2016
  • FANCJ is a DNA helicase which contributes genome stability by resolving G-quadruplex DNA from 5' to 3' direction. In addition to main ATPase helicase core, FANCJ has the protein binding region at its C-terminal part. BRCA1 and BLM are the binding partner of FANCJ and these protein-protein interactions contribute genomic stability and the proper response to replication stress. As the first attempt for studying FANCJ-BLM interaction, we prepared BLM binding region of FANCJ and characterized with CD and NMR spectroscopy. FANCJ (881-941) with N-ter 6xHis was purified as the oligomer. Secondary structure prediction based on CD data revealed that FANCJ (881-941) composed with ${\beta}$ sheet, turn and coils.$^1H-^{15}N$ HSQC spectra showed nonhomogeneous peak intensities with less number of peaks comparing than the number of amino acids in the construct. It indicated that optimization should be necessary for detailed further structural studies.

Acute hepatic injury following ischemia and reperfusion in rats

  • Park, Mee-Jung-;Lee, Sang-Ho-;Park, Doo-Soon-;Cho, Tai-Soon;Lee, Sun-Mee-
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.340-340
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    • 1994
  • Since total hepatic ischemia(IS) occurs with transplantation, there has been interest in evaluating hepatic function after ischemia and subsequent reflow of blood. Four groups of animals were studied: group 1 (sham), group 2 (30mins IS), group 3 (60mins IS), and g.cup 4 (90mins IS). Serum transaminase(STA), wet weight-to-dry weight ratio(W/D), lipid peroxides(LPO), glucose-6-phosphatase(G-6-Pase) activity, Na$\^$+//K$\^$+/-ATPase(ATPase) activity were measured at 1, 5 and 24hrs after hepatic ischemia. Significant changes occurred between 1 and 5hrs of reperfusion. STA was 3579${\pm}$401, 4593${\pm}$675 and 6348${\pm}$808 U/L in group 2, 3 and 4 respectively. These changes were ischemic time-dependent manner. W/D in group 3 and 4 were significantly increased than that in sham group at all time points measured. In sham group, the level of LPO in the liver microsome remained constant at approximately 0. 5nmole MDA formed/mg protein througllout the experiment, In all ischemic groups on the other hand, the level of LPO started to increase at ischemia and markedly increased at all reperfusion period. Similar to STA, these changes were also dependent on duration of ischemia. Although G-6-Pase activity remained unchanged in both group 2 and group 3 until 5hrs of reperfusion, marked decrease in G-6-Pase activity was observed at grcup 4. ATPase activity was significantly decreased at 1, 5 and 24 hrs of reperfusion in group 3, whereas it was not changed in group 2. Furthermore, ATPase activity in group 4 started to decrease at ischemia and markedly decreased for entire reperfusion period. These data suggest that severity of hepatocellular injury is associated with period of ischemia as well as period of reperfusion.

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Characterizing Salt Stress Response in a Rice Variety and Its Salt Tolerant Lines Derived from In Vitro Mutagenesis

  • Lee In Sok;Kim Dong Sub;Kang Si Yong;Wi Seung Gon;Jin Hua;Yun PiI-Yong;Lim Yong Pyo;Lee Young Il
    • Journal of Plant Biotechnology
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    • v.6 no.4
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    • pp.205-212
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    • 2004
  • The objectives were to compare the salt tolerance levels in the parental rice cultivar, Dongjinbyeo, and induced mutagenesis derived its lines for plant height, MDA, ATPase, POD, and 2-dimensional protein electrophoresis pattern in NaCl-containing hydroponic nutrient solutions. Rice plants isolated from a population of rice (Oryza sativa L. cv. Dongjinbyeo) mutation lines, which were generated in combination with in vitro selection and gamma-ray, exhibited salt tolerance. Line No. 18 had the longest plant, whereas NaCl-sensitive line (No. 25) had the shortest plant. The parent, and the sensitive line showed severe damage from salt stress. Tolerant lines (No. 18, 50) had a lower malonaldehyde (MDA) content than the sensitive one (Dongjinbyeo, No. 25) during salt stress. Several proteins showed significant quantitative variation through 2DE; phosphoribulokinase, peroxidase, oxygen evolving enhancer 1 and the $H^+-ATPase$, which are known to be involved in salt tolerance. The effect of salt on peroxidase and $H^+-ATPase$ activity in the seedlings of two groups with contrasting genotypes of rice was studied. A greater activity was recorded in the tolerant lines as compared to the sensitive ones (P<0.05, Duncan's test). The results indicate that salt tolerant lines expressed more salt stress-inducible proteins associated with salt tolerance than the sensitive lines during salt stress.

$Na^{+}$ Metabolism in the Frog in the Winter Time (겨울철 개구리 (Rana temporaria)의 $Na^{+}$이온 대사)

  • Lee, Joong-Woo;Kang, Doo-Hee
    • The Korean Journal of Physiology
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    • v.7 no.1
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    • pp.41-47
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    • 1973
  • $Na^{+}$ balance was studied in Rana temporaria, which hibenates in fresh water in the winter time. $Na^{+}$ uptake rate, skin $Na^{+}$ loss rate, urinary $Na^{+}$ loss rate and $Na^{+}-K^{+}$ adenosine triphosphatase(ATPase) were measured at two different temperatures $1{\sim}2^{\circ}C\;and\;20{\sim}24^{\circ}C$ respectively. The results obtained were as follows: 1. $Na^{+}$ uptake rates in the frog in an artificial Pond water (APW) were found to be $8.28{\pm}0.73\;and\;2.19{\pm}0.37\;{\mu}Eq/g/day\;at\;20{\sim}24^{\circ}C\;and\;1.0{\sim}2.5^{\circ}$ respectively. 2. $Na^{+}$ loss rate through the frog skin to APW were found to be $4.26{\pm}0.72\;and\;0.93{\pm}0.21\;{\mu}Eq/g/day$ at the same temperatures. 3. Mean rates of urinary $Na^{+}$ loss at $20{\sim}24^{\circ}C\;and\;3{\sim}4^{\circ}C$ were found to be $3.02{\pm}0.73\;and\;0.78{\pm}0.13\;{\mu}Eq/g/day$ respectively. 4. The activities of $Na^{+}-K^{+}$ activated ATPase of frog skin fragments were found to be $258{\pm}39.4\;and\;49.6{\pm}7.1\;{\mu}M\;Pi/g$ protein/hr at $24^{\circ}C\;and\;2^{\circ}C$ respectively. From the above results, it may be concluded that frogs can take up enough $Na^{+}$ through the skin from APW exceeding skin loss Plus urinary loss at $1{\sim}2^{\circ}C$. It is suggested that $Na^{+}$ transport across frog skin is closely related with $Na^+-K^+$ ATPase since $Q_{10}\;of\;Na^{+}$ uptake is much similar to that of the activities of $Na^{+}-K^{+}$ ATPase.

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Up-Regulation of RANK Expression via ERK1/2 by Insulin Contributes to the Enhancement of Osteoclast Differentiation

  • Oh, Ju Hee;Lee, Na Kyung
    • Molecules and Cells
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    • v.40 no.5
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    • pp.371-377
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    • 2017
  • Despite the importance of the receptor activator of nuclear factor (NF)-kappaB ligand (RANKL)-RANK signaling mechanisms on osteoclast differentiation, little has been studied on how RANK expression is regulated or what regulates its expression during osteoclastogenesis. We show here that insulin signaling increases RANK expression, thus enhancing osteoclast differentiation by RANKL. Insulin stimulation induced RANK gene expression in time- and dose-dependent manners and insulin receptor shRNA completely abolished RANK expression induced by insulin in bone marrow-derived monocyte/macrophage cells (BMMs). Moreover, the addition of insulin in the presence of RANKL promoted RANK expression. The ability of insulin to regulate RANK expression depends on extracellular signal-regulated kinase 1/2 (ERK1/2) since only PD98059, an ERK1/2 inhibitor, specifically inhibited its expression by insulin. However, the RANK expression by RANKL was blocked by all three mitogen-activated protein (MAP) kinases inhibitors. The activation of RANK increased differentiation of BMMs into tartrate-resistant acid phosphatase-positive ($TRAP^+$) osteoclasts as well as the expression of dendritic cell-specific transmembrane protein (DC-STAMP) and d2 isoform of vacuolar ($H^+$) ATPase (v-ATPase) Vo domain (Atp6v0d2), genes critical for osteoclastic cell-cell fusion. Collectively, these results suggest that insulin induces RANK expression via ERK1/2, which contributes to the enhancement of osteoclast differentiation.