• Title/Summary/Keyword: SR protein

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Cyclosporine A (Cipol-$N^{(R)}$) Therapy in Children with Idiopathic Nephrotic Syndrome (소아 특발성 신증후군에서의 cyclosporine A(Cipol-$N^{(R)}$)의 치료 효과)

  • Hong Ihn Hee;Ko Cheol Woo;Koo Ja Hoon;Kim Ji-Hong;Kim Pyung-Kil;Cho Byoung Soo
    • Childhood Kidney Diseases
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    • v.3 no.1
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    • pp.48-56
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    • 1999
  • Purpose : This multicenter collaboratory study was conducted to see the therapeutic efficacy and side effect of cyclosporine A (Cipol-$N^{(R)}$, Chong Kun Dang) on children with idiopathic nephrotic syndrome who experienced frequently relapsing (FR), steroid dependent (SD), or steroid resistant (SR) pattern. Patients and methods : Thirty-nine children with SD/FR NS and 3 children with SR NS were enrolled in the study. After induction of remission (SD/FR NS) with steroid or after 4 weeks of steroid therapy (SR NS), cyclosporine A was started in a dose of 4-5 mg/Kg/day in two divided dose and steroid (prednisolone or equivalent dose of deflazacort) was tapered slowly. During 16 weeks of study period, monthly check up of physical examination and various laboratory tests including BUN, creatinine, Ccr and cyclosporine blood level were done. Results : Out of 39 children with SD/FR NS, 35($89.7\%$) maintained sustained remission and at 4 weeks after therapy, values of serum protein, albumin, cholesterol, and 24 hours urinary protein excretion showed normal values. Two out of 3 children with SR NS showed and sustained remission with cyclosporine A therapy. Side reaction to cyclosporine A therapy showed hypertrichosis in 8 cases and hyperuricemia in 5 cases. However, other laboratory tests including CBC, liver profile, BUN, creatinine and GFR (creatinine clearance utilizing 24 hour urine) did not show any abnormalities during the 16 weeks of study period. Conclusion : Cyclosporine A (Cipoi-$N^{(R)}$ Chong Kun Dang) can be utilized quite effectively on children with SD/FR or SR NS and further trial of cyclosporine A on long-term basis (1-2 year period) is needed to determine it's efficacy and side effect (especially nephrotoxicity) of long-term administration of cyclosporine A.

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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.

The Role of Actin Binding Protein -Caldesmon- of the Mechanism of $Ca^{2+}$-dependent/-independent Smooth Muscle Contraction - Approach of Basic Medical for the Study of Senile Cardiovascular Disease-related Senile Physical Therapy - (세포 내 $Ca^{2+}$-의존성/-비의존성 평활근 수축기전에 대한 액틴결합단백질-Caldesmon-의 역할 - 노인성 심혈관질환 관련 노인물리치료 연구를 위한 기초의학적 접근 -)

  • Kim, Jung-Hwan;Min, Kyung-Ok;Choi, Young-Duk;Lee, Joon-Hee;Chon, Ki-Young
    • Journal of Korean Physical Therapy Science
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    • v.11 no.1
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    • pp.20-27
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    • 2004
  • It is widely accepted that smooth muscle contraction is triggered by intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) released from intracellular $Ca^{2+}$ stores such as sarcoplasmic reticulum (SR) and from the extracellular space, The increased $[Ca^{2+}]_i$ can phosphorylate the 20-kDa myosin light chain ($MLC_{20}$) by activating MLC kinase (MLCK), and this initiates smooth muscle contraction. In addition to the $[Ca^{2+}]_i$-MLCK-tension pathway, a number of intracellular signal molecules, including mitogen-activated protein kinase (MAPK), protein kinase C (PKC), phosphatidylinositol 3-kinase (PI3K), and Rho-associated coiled coil-forming protein kinase (ROCK), play important roles in the regulation of smooth muscle contraction. However, the mechanisms regulating contraction of caldesmon (CaD), actin-binding protein, are not entirely elucidated in the presence of $Ca^{2+}$. It is known that CaD tightly interacts with actin and inhibits actomyosin ATPase activity. Therefore, the purpose of the present study was to investigate the roles of $Ca^{2+}$-dependent CaD in smooth muscle contraction. Endothelin-1 (ET-1), G-protein coupled receptor agonist and vasoconstrictor, increased both vascular smooth contraction and phosphorylation of CaD in the presence of $Ca^{2+}$. These results suggest that ET-1 induces contraction and phosphorylation of CaD in rat aortic smooth muscle, which may he mediated by the increase of $[Ca^{2+}]_i$.

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Effects of 『Geum-Gwe-Yo-Ryak(金匱要略)』 Prescription for Chest Pain Including Kwaruhaebaekbanha-tang and Kwaruhaebaekpaekju-tang on Macrophage Polarization (금궤요략(金匱要略) 심통 처방 중 과루해백반하탕과 과루해백백주탕이 대식세포 극성화에 미치는 영향)

  • Son, Chang-Hyeon;Lee, Sang-Min;Yu, Ga-Ram;Lee, Seung-Jun;Lim, Dong-Woo;Kim, Hyuck;Park, Won-Hwan;Kim, Jai-Eun
    • The Journal of Korean Medicine
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    • v.40 no.2
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    • pp.51-62
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    • 2019
  • Objectives: This study was designed to evaluate the macrophages polarization of traditional Korean medicine on cardiac pain about Geum-Gwe-Yo-Ryak's two prescriptions including Kwaruhaebaekbanha-tang (KHB) and Kwaruhaebaekpaekju-tang (KHP). Materials and methods: Flow cytometry analysis was used to measure the changes in the ratio of M1 type and M2 type macrophages. Protein expression of nuclear factor-like 2 (Nrf2), heme oxygenase-1 (HO-1), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) were measured by Western Blot, and ABCA1 and SR-B1 were detected by real time PCR (RT-PCR). Intracellular lipid accumulation was measured by Oil Red O staining (ORO staining). Results: KHB and KHP increase anti-oxidative activity related protein levels including Nrf2 and HO-1. Furthermore, KHB and KHP inhibit lipid accumulation on intracellular levels through induction of ATP binding receptor cassette subfamily A member 1 (ABCA1) and scavenging receptor class B member 1 (SR-B1), respectively. Finally, KHB and KHP also blocked pro-inflammatory mediators including tumor necrosis factor-alpha ($TNF{\alpha}$) and interleukin-6 (IL-6), iNOS and COX-2 expression. Conclusion: This study suggests that KHB and KHP potently regulate the M1/M2 macrophage polarization.

Identification of binding motifs for skeletal ryanodine receptor and triadin

  • Lee, Jae-Man;Kim, Do-Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.66-66
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    • 2003
  • In skeletal muscle cells, depolarization of the transverse tubules (T-tubules) results in Ca$\^$2+/ release from the sarcoplasmic reticulum (SR), leading to elevated cytoplasmic Ca$\^$2+/ and muscle contraction. This process has been known as excitation-contraction coupling (E-C coupling). Several proteins, such as the ryanodine receptor (RyR), triadin, junctin, and calsequestrin (CSQ), have been identified to be involved in the Ca$\^$2+/ release process. However, the molecular interactions between the SR proteins have not been resolved. In the present study, the mechanisms of interaction between RyRl and triadin have been studied by in vitro protein binding and $\^$45/Ca$\^$2+/ overlay assays. Our data demonstrate that the intraluminal loop II of RyR1 binds to triadin in Ca$\^$2+/-independent manner. Moreover, we could not find any Ca$\^$2+/ binding sites in the loop II region. GST-pull down assay revealed that a KEKE motif of triadin, which was previously identified as a CSQ binding site (Kobayasi et al.,2000 JBC) was also a binding site for RyR1. Our results suggest that the intraluminal loop II of RyR could participate in the RyR-mediated Ca$\^$2+/ release process by offering a direct binding site to luminal triadin.

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Cardiac hypertrophy and abnormal $Ca^{2+}$ handling in transgenic mice overexpressing jnnctate

  • Hong, Chang-Soo;Cho, Myeong-Chan;Kwak, Yong-Geun;Chane, Soo-Wan;Kim, Do-Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.52-52
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    • 2003
  • Junctate is a newly identified integral ER/SR membrane $Ca^{2+}$ binding protein, which is an alternative splicing form of the same gene generating aspartyl $\square$-hydroxylase and junctin. To elucidate the functional role of junctate in heart, transgenic (TG) mice overexpressing mouse cardiac junctate-1 under the control of mouse $\square$$^{~}$ myosin heavy chain promoter were generated. Overexpression of junctate in mouse heart resulted in cardiac hypertrophy, increased fibrosis, bradycardia, arrhythmias and impaired contractility. Overexpression of junctate also led to down-regulation of SERCA2, calsequestrin, calreticulin and RyR, but to up-regulation of NCX and PMCA. The SR $Ca^{2+}$ content decreased and the L-type $Ca^{2+}$ current density and the action potential durations increased in TG cardiomyocytes, which could be the cause for the bradycardia in TG heart. The present work has provided an important example of pathogenesis leading to cardiac hypertrophy and arrhythmia, which was caused by impaired $Ca^{2+}$ handling by overexpression of junctate in heart.n heart.

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Identification of B52-dependent Gene Expression Signature and Alternative Splicing Using a D. melanogaster B52-null Mutant

  • Hong, Sun-Woo;Jung, Mi-Sun;Kim, Eun-Kyung;Lee, Dong-Ki;Kim, So-Youn
    • Bulletin of the Korean Chemical Society
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    • v.30 no.2
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    • pp.323-326
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    • 2009
  • SR proteins are essential splicing regulators and also modulate alternative splicing events, which function both as redundant and substrate-specific manner. The Drosophila B52/SRp55, a member of the SR protein family, is essential for the fly development in vivo, as deletion of B52 gene results in lethality of animals at the second instar larval stage. Identification of the splicing target genes of B52 thus should be crucial for the understanding of the specific developmental role of B52 in vivo. In this study, we performed whole-genome DNA microarray experiments with a B52- knock-out animal. Analysis of the microarray data not only provided the B52-dependent gene expression signature, but also revealed a larval-stage specific, alternative splicing target gene of B52. Our result thus provides a starting point to understand the essential function of B52 at the organismal level.

Detection of Insect Pathogen Serratia marcescens in Protaetia brevitarsis seulensis (Kolbe) from Korea

  • Kwak, Kyu-Won;Han, Myung-Sae;Nam, Sung-Hee;Choi, Ji-Young;Lee, Seok-Hyun;Choi, Young-Cheol;Park, Kwan-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.28 no.2
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    • pp.25-31
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    • 2014
  • Protaetia brevitarsis seulensis (Kolbe) is widely used in Korea, as a protein-rich, alternate, functional food with pharmacological benefits. In addition to anti-oxidant properties, the larvae of P. b. seulensis also show positive effects against hepatic disorder and diabetes; therefore, P. b. seulensis larvae are being reared on a large scale in Korea. We evaluated reared larvae of P. b. seulensis from Gyeong-gi in Korea. Using 16SrRNA PCR, electro-microscopy, and bioassay techniques, we found that the larvae harbored Spo-1, a bacterium identified as the insect pathogen Serratia marcescens. Therefore, we highlight the use of this insect as an alternate food and the need for its sanitary rearing conditions, as contamination may affect public health.

Investigation of the effect of SRSF9 overexpression on HIV-1 production

  • Ga-Na, Kim;Kyung-Lee, Yu;Hae-In, Kim;Ji Chang, You
    • BMB Reports
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    • v.55 no.12
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    • pp.639-644
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    • 2022
  • Serine-arginine-rich splicing factors (SRSFs) are members of RNA processing proteins in the serine-arginine-rich (SR) family that could regulate the alternative splicing of the human immunodeficiency virus-1 (HIV-1). Whether SRSF9 has any effect on HIV-1 regulation requires elucidation. Here, we report for the first time the effects and mechanisms of SRSF9 on HIV-1 regulation. The overexpression of SRSF9 inhibits viral production and infectivity in both HEK293T and MT-4 cells. Deletion analysis of SRSF9 determined that the RNA regulation motif domain of SRSF9 is important for anti-HIV-1 effects. Furthermore, overexpression of SRSF9 increases multiple spliced forms of viral mRNA, such as Vpr mRNA. These data suggest that SRSF9 overexpression inhibits HIV-1 production by inducing the imbalanced HIV-1 mRNA splicing that could be exploited further for a novel HIV-1 therapeutic molecule.

Analysis of Dry Matter Yield and Feed Value for Selecting of Whole Crop Rice (최적 총체사료벼 품종 선발을 위한 건물수량 및 사료가치 분석)

  • Lee, Jeom-Ho;Jeong, O.Y.;Paek, J.S.;Hong, H.C.;Yang, S.J.;Lee, Y.T.;Kim, J.G.;Sung, K.I.;Kim, B.W.
    • Journal of Animal Science and Technology
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    • v.47 no.3
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    • pp.355-362
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    • 2005
  • This study was carried out to obtain basic information on variety selection for the utilization of whole crop rices(WCR) at National Institute of Crop Science, RDA, in 2004. Fifteen varieties and elite line were evaluated on feed value such as dry matter yield(DMY), crude protein( CP), acid detergent fiber(ADF), neutral detergent fiber(NDF) and total digestible nutrients(TDN). The dry matter yields were ranged from 13.23 to 17.83 ton per ha, the highest yielding varieties were Sobibyeo(l6.98ton / ha) in Japonica type, SR22060 (17.83 ton / ha) in New plant type, Hangangchalbyeo(I7.66 ton / ha) in Tongil type. Suweon 468 showed the highest value in the RFV and TDN content among the varieties, and Suweon 468, Suweon 498, Suweon 490 and SR22058 were chosen to have the high feed values through cluster analysis. The dry weight(grain) was found to be positively related with percent of the ripened grain, 1,000 grain weight and CPo TDN content was found to be positively related with CP, but negatively related with NDF and ADF. RFV was found to be negatively related with plant height, NDF and ADF. The promising rice varieties for WCR were Suweon 468, Suweon 498, Suweon 490 and SR22058 on the basis of CP, TDN and DMY.