• Title/Summary/Keyword: RyR

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Structural characterization of calmodulin like domain of ryanodine receptor type 1

  • Song, Yonghyun;Kang, Sunmi;Park, Sunghyouk
    • Journal of the Korean Magnetic Resonance Society
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    • v.19 no.2
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    • pp.74-82
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    • 2015
  • Ryanodine receptor (RyR) is one of the two major $Ca^{2+}$ channels in membranes of intracellular $Ca^{2+}$ stores and is found in sarcoplasmic reticulum (SR), endoplasmic reticulum (ER). RyR1 is also the major calmodulin-binding protein of sarcoplasmic reticulum membranes. Residues 4064-4210 in the RyR1 polypeptide chain has similar primary sequence with calmodulin (CaM) and was designated as CaM-like domain (CaMLD). When expressed as a recombinant peptide, CaMLD showed several CaM-like properties in previous studies. Still, previous studies of CaMLD were focused on protein-protein interactions rather than its own properties. Here, we studied the expression of CaMLD and its sub-domains corresponding to each lobe of CaM in Escherichia coli. CaMLD could be obtained only as inclusion body, and it was refolded using urea solubilization followed by dialysis. Using spectroscopic approaches, such as NMR, circular dichroism, and gel filtration experiment, we found that the refolded CaMLD exists as nonspecific aggregate, even though it has alpha helical secondary structure. In comparison, the first half of CaMLD (R4061-4141) could be obtained as natively soluble protein with thioredoxin fusion. After the removal of the fusion tag, it exhibited folded and helical properties as shown by NMR and circular dichroism experiments. Its oligomeric status was different from CaMLD, existing as dimeric form in solution. However, the second half of the protein could not be obtained as soluble protein regardless of fusion tag. Based on these results, we believe that CaMLD, although similar to CaM in sequence, has quite different physicochemical properties and that the second half of the protein renders it the aggregative properties.

Skeletal Ryanodine Receptor 1-Heterozygous PSE (Pale, Soft and Exudative) Meat Contains a Higher Concentration of Myoglobin than Genetically Normal PSE Meat in Pigs

  • Obi, T.;Matsumoto, M.;Miyazaki, K.;Kitsutaka, K.;Tamaki, M.;Takase, K.;Miyamoto, A.;Oka, T.;Kawamoto, Y.;Nakada, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.9
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    • pp.1244-1249
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    • 2010
  • Comparisons of properties between skeletal ryanodine receptor 1 (sRyR1)-heterozygous-mutated and normal types of meat were carried out in pigs using PSE (pale, soft and exudative) meat found during the butchering process. All samples considered to be PSE meat showed irregular running and disorder of the muscle fibers and a wider inter-fiber space upon light microscopic observation. Electron microscopy revealed disintegration, twisting, and disorder of the myofibril arrangement and elimination of the Z line in PSE meat, compared with normal meat. Meat property tests demonstrated greater decreases in water holding capacity, moisture and sarcoplasmic protein, and higher $L^*$ values for the meat color index in PSE meat than in normal meat, but there were no differences in these factors between genetically normal and sRyR1-heterozygous PSE meat. On the other hand, higher $a^*$ and $b^*$ values were observed in sRyR1-heterozygous than in normal PSE meat, and similar alterations to the a* value were observed in terms of the amount of myoglobin and density of the 17-kDa protein band, corresponding to the molecular mass of myoglobin, on SDS-PAGE gels. These results suggest that sRyR1-heterozygous PSE pork contains much more myoglobin than genetically normal PSE meat.

Alteration of Ryanodine-receptors in Cultured Rat Aortic Smooth Muscle Cells

  • Kim, Eun-Ji;Kim, Dong-Kwan;Kim, Shin-Hye;Lee, Kyung-Moo;Park, Hyung-Seo;Kim, Se-Hoon
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.6
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    • pp.431-436
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    • 2011
  • Vascular smooth muscle cells can obtain a proliferative function in environments such as atherosclerosis in vivo or primary culture in vitro. Proliferation of vascular smooth muscle cells is accompanied by changes in ryanodine receptors (RyRs). In several studies, the cytosolic $Ca^{2+}$ response to caffeine is decreased during smooth muscle cell culture. Although caffeine is commonly used to investigate RyR function because it is difficult to measure $Ca^{2+}$ release from the sarcoplasmic reticulum (SR) directly, caffeine has additional off-target effects, including blocking inositol trisphosphate receptors and store-operated $Ca^{2+}$ entry. Using freshly dissociated rat aortic smooth muscle cells (RASMCs) and cultured RASMCs, we sought to provide direct evidence for the operation of RyRs through the $Ca^{2+}$- induced $Ca^{2+}$ -release pathway by directly measuring $Ca^{2+}$ release from SR in permeabilized cells. An additional goal was to elucidate alterations of RyRs that occurred during culture. Perfusion of permeabilized, freshly dissociated RASMCs with $Ca^{2+}$ stimulated $Ca^{2+}$ release from the SR. Caffeine and ryanodine also induced $Ca^{2+}$ release from the SR in dissociated RASMCs. In contrast, ryanodine, caffeine and $Ca^{2+}$ failed to trigger $Ca^{2+}$ release in cultured RASMCs. These results are consistent with results obtained by immunocytochemistry, which showed that RyRs were expressed in dissociated RASMCs, but not in cultured RASMCs. This study is the first to demonstrate $Ca^{2+}$ release from the SR by cytosolic $Ca^{2+}$ elevation in vascular smooth muscle cells, and also supports previous studies on the alterations of RyRs in vascular smooth muscle cells associated with culture.

Capacity of Activated Carbon Derived from Agricultural Waste in the Removal of Reactive Dyes from Aqueous Solutions

  • Manoochehri, Mahboobeh;Rattan, V.K.;Khorsand, Ameneh;Panahi, Homayon Ahmad
    • Carbon letters
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    • v.11 no.3
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    • pp.169-175
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    • 2010
  • The study describes the results of batch experiments on the removal of Reactive Yellow 15 (RY15) and Reactive Black 5 (RB5) from synthetic textile wastewater onto Activated Carbon from Walnut shell (ACW). The experimental data were analyzed by the Langmuir, Freundlish, Temkin and Dubinin-Radushkevich (D-R) models of adsorption. The experiments were carried out as function of initial concentrations, pH, temperature (303-333), adsorbent dose and kinetics. The surface area and pore volumes of adsorbent were measured by BET and BJH methods. The findings confirm the surface area (BET) is 248.99 $m^2/g$. The data fitted well with the Temkin and D-R isotherms for RY15 and RB5, respectively. The most favorable adsorption occurred in acidic pH. Pseudo-second order kinetic model were best in agreement with adsorption of RY15 and RB5 on ACW. The results indicate that walnut shell could be an alternative to more costly adsorbent currently being used for dyes removal.

Characterization of calumenin in mouse heart

  • Sahoo, Sanjaya Kumar;Kim, Do-Han
    • BMB Reports
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    • v.43 no.3
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    • pp.158-163
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    • 2010
  • Calumenin is a multiple EF-hand $Ca^{2+}$-binding protein located in the endo/sarcoplasmic reticulum of mammalian hearts. Calumenin belongs to the CREC family of $Ca^{2+}$-binding proteins having multiple EF-hands. $Ca^{2+}$ homeostasis in the sarcoplasmic reticulum (SR) of mammalian hearts is maintained by RyR2, SERCA2 and other associated SR resident proteins. Evidence suggests that calumenin interacts with RyR2 and SERCA2, and therefore changes in the expression of calumenin could alter $Ca^{2+}$ cycling in mouse heart. In this review, current knowledge of the biochemical and functional roles of calumenin in mouse heart is described.

Effects of Recombinant Imperatoxin A $(IpTx_a$ mutants on $Ca^{2+}$ Release Channel/Ryanodine Receptor in Rabbit Skeletal Sarcoplasmic Reticulum

  • Seo, In-Ra;Park, Murim;Kim, Do-Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 1999.06a
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    • pp.55-55
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    • 1999
  • Imperatoxin A (IpTx$_{a}$), a 3.7 kDa peptide from the African scorpion Pandinus imperator, has been known as an agonist of skeletal ryanodine receptor (RyR). In order to study the structure and function of the toxins on RyR, the IpTx$_{a}$ cDNA was PCR-amplified using 3 pairs of primers and the toxin was expressed in E. coli expression system.(omitted)ted)

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Interplay Between Intra- and Extracellular Calcium Ions

  • Lee, Eun Hui;Kim, Do Han;Allen, Paul D.
    • Molecules and Cells
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    • v.21 no.3
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    • pp.315-329
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    • 2006
  • Two, well characterized cationic channels, the ryanodine receptor (RyR) and the canonical transient receptor potential cation channel (TRPC) are briefly reviewed with a particular attention on recent developments related to the interplay between the two channel families.

The Production Objectives and Optimal Standard of Density Control Using Stand Density Management Diagram for Pinus densiflora Forests in Korea (임분밀도관리도를 이용한 소나무림의 적정 임분밀도 관리 기준 및 수확목표)

  • Park, Joon-hyung;Jung, Su-Young;Yoo, Byung-oh;Lee, Kwang-Soo;Park, Yong-bae;Kim, Hyung-Ho
    • Journal of Korean Society of Forest Science
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    • v.106 no.4
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    • pp.457-464
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    • 2017
  • This study has utilized the stand density management diagram to devise an efficient management standard for the stand density for Pinus densiflora that secures the health of the stands and predicted the harvest goals. The appropriate stand control level was estimated by modeling the relationship of the relative yield index (Ry) to the ratio of slender trees within the stand through an exponential function; the coefficient of determination ($R^2$) was found to be 0.424 according to the estimation. The ratio of slender trees within the stand showed a tendency of rapid increase at a certain relative yield index; with this relational function, the appropriate Ry value of 0.84 was obtained. By estimating the curve of the Ry value 0.84, which was the appropriate stand density management level, as well as the height of dominant trees in the central region of Korea, the production objective for each site index was set. Assuming that the final age by the site indices ranged from 10 to 16 for the P. densiflora in central region of Korea, the number of production was estimated to be between 426 to 1,311 trees per ha. It was predicted that the production of medium-diameter logs larger than 30 cm in diameter is possible for the target DBH at a site index of more than 16; small-diameter logs larger than 20 cm in diameter for site indices 12 and 14 enabled, and small-diameter logs of less than 20 cm for site index 10.

Unaided Stapling Technique for Pure Single-Incision Distal Gastrectomy in Early Gastric Cancer: Unaided Delta-Shaped Anastomosis and Uncut Roux-en-Y Anastomosis

  • Suh, Yun-Suhk;Park, Ji-Ho;Kim, Tae Han;Huh, Yeon-Ju;Son, Young Gil;Yang, Jun-Young;Kong, Seong-Ho;Lee, Hyuk-Joon;Yang, Han-Kwang
    • Journal of Gastric Cancer
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    • v.15 no.2
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    • pp.105-112
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    • 2015
  • Purpose: Intracorporeal anastomosis is the most difficult procedure during pure single-incision distal gastrectomy (SIDG) that affects its generalization. We introduced unaided delta-shaped anastomosis (uDelta), a novel anastomosis technique, for gastroduodenostomy after pure SIDG, and compared the results with those of previously reported Roux-en-Y anastomosis (RY). Materials and Methods: Between March 2014 and March 2015, SIDG with D1+ lymph node dissection was performed for early gastric cancer through a 2.5-cm transumbilical incision without any additional port. uDelta was performed by the operator alone, without any intracorporeal assistance. Results: uDelta was performed on 11 patents, and uncut RY was performed on 5-patients without open or multiport conversion. R0 resection was performed in all cases. No significant differences were observed in mean age and body mass index between patients who underwent uDelta or RY. Mean operation times were $214.5{\pm}36.2$ minutes for uDelta and $240.8{\pm}65.9$ minutes for RY, which was not significantly different. Reconstruction time for uDelta was shorter than that for RY, with marginal statistical significance ($26.1{\pm}8.3$ minutes vs. $38.0{\pm}9.1$ minutes, P=0.05). There were no intraoperative transfusions, 30-day mortality, or anastomosis-related complications in either group. Average length of hospital stay was $8.2{\pm}1.9$ days in the uDelta group and $7.2{\pm}0.8$ days in the RY group (P=0.320). Conclusions: After carefully considering indications, uDelta can be a feasible and can be a reproducible reconstruction method after SIDG in early gastric cancer.

Ryanodine Receptor-mediated Calcium Release Regulates Neuronal Excitability in Rat Spinal Substantia Gelatinosa Neurons

  • Park, Areum;Chun, Sang Woo
    • International Journal of Oral Biology
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    • v.40 no.4
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    • pp.211-216
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    • 2015
  • Nitric Oxide (NO) is an important signaling molecule in the nociceptive process. Our previous study suggested that high concentrations of sodium nitroprusside (SNP), a NO donor, induce a membrane hyperpolarization and outward current through large conductances calcium-activated potassium ($BK_{ca}$) channels in substantia gelatinosa (SG) neurons. In this study, patch clamp recording in spinal slices was used to investigate the sources of $Ca^{2+}$ that induces $Ca^{2+}$-activated potassium currents. Application of SNP induced a membrane hyperpolarization, which was significantly inhibited by hemoglobin and 2-(4-carboxyphenyl) -4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (c-PTIO), NO scavengers. SNP-induced hyperpolarization was decreased in the presence of charybdotoxin, a selective $BK_{Ca}$ channel blocker. In addition, SNP-induced response was significantly blocked by pretreatment of thapsigargin which can remove $Ca^{2+}$ in endoplasmic reticulum, and decreased by pretreatment of dentrolene, a ryanodine receptors (RyR) blocker. These data suggested that NO induces a membrane hyperpolarization through $BK_{ca}$ channels, which are activated by intracellular $Ca^{2+}$ increase via activation of RyR of $Ca^{2+}$ stores.