• 제목/요약/키워드: Protein Stabilization

검색결과 145건 처리시간 0.024초

RNA Binding Protein Rbms1 Enables Neuronal Differentiation and Radial Migration during Neocortical Development by Binding and Stabilizing the RNA Message for Efr3a

  • Habib, Khadija;Bishayee, Kausik;Kang, Jieun;Sadra, Ali;Huh, Sung-Oh
    • Molecules and Cells
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    • 제45권8호
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    • pp.588-602
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    • 2022
  • Various RNA-binding proteins (RBPs) are key components in RNA metabolism and contribute to several neurodevelopmental disorders. To date, only a few of such RBPs have been characterized for their roles in neocortex development. Here, we show that the RBP, Rbms1, is required for radial migration, polarization and differentiation of neuronal progenitors to neurons in the neocortex development. Rbms1 expression is highest in the early development in the developing cortex, with its expression gradually diminishing from embryonic day 13.5 (E13.5) to postnatal day 0 (P0). From in utero electroporation (IUE) experiments when Rbms1 levels are knocked down in neuronal progenitors, their transition from multipolar to bipolar state is delayed and this is accompanied by a delay in radial migration of these cells. Reduced Rbms1 levels in vivo also reduces differentiation as evidenced by a decrease in levels of several differentiation markers, meanwhile having no significant effects on proliferation and cell cycle rates of these cells. As an RNA binding protein, we profiled the RNA binders of Rbms1 by a cross-linked-RIP sequencing assay, followed by quantitative real-time polymerase chain reaction verification and showed that Rbms1 binds and stabilizes the mRNA for Efr3a, a signaling adapter protein. We also demonstrate that ectopic Efr3a can recover the cells from the migration defects due to loss of Rbms1, both in vivo and in vitro migration assays with cultured cells. These imply that one of the functions of Rbms1 involves the stabilization of Efr3a RNA message, required for migration and maturation of neuronal progenitors in radial migration in the developing neocortex.

나노 입자의 피커링 안정화를 이용한 식품 에멀젼의 최근 연구동향 (A Recent Research Trends for Food Emulsions using Pickering Stabilization of Nano-particles)

  • 조완구
    • 한국응용과학기술학회지
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    • 제29권2호
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    • pp.238-247
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    • 2012
  • 식품산업에서의 나노기술은 많은 연구와 가능성 있는 제품으로 새롭게 각광받는 분야이다. 나노와 마이크로 크기의 고체 입자는 유화된 오일 또는 물 입자를 함유하는 분산계의 처방에 적용 가능성이 매우 높은 것으로 인식되고 있다. 본 논문은 식용의 지방 결정, 실리카 나노 입자 및 생체 유래의 나노 결정체에 기반을 둔 식품 에멀젼의 성질과 개발에 관한 리뷰논문으로 다양한 피커링 안정화 분야에 관하여 검토하였다. 특히, 단백질로 안정화된 일반적인 에멀젼과 비교하여 Pickering 안정화 입자에 대해 초점을 맞추고자한다. 또한, 식품 산업에서 이들의 응용 예를 다루고자한다.

Immobilization and Stability of Lipase from Mucor racemosus NRRL 3631

  • Adham, Nehad Zaki;Ahmed, Hanan Mostafa;Naim, Nadia
    • Journal of Microbiology and Biotechnology
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    • 제20권2호
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    • pp.332-339
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    • 2010
  • The lipase from Mucor racemosus NRRL 3631 was partially purified by fractional precipitation using 60% ammonium sulfate, which resulted in a 8.33-fold purification. The partially purified lipase was then immobilized using different immobilization techniques: physical adsorption, ionic binding, and entrapment. Entrapment in a 4% agar proved to be the most suitable technique (82% yield), as the immobilized lipase was more stable at acidic and alkaline pHs than the free enzyme, plus 100% of the original activity was retained owing to the thermal stability of the immobilized enzyme after heat treatment for 60 min at $45^{\circ}C$. The calculated half-lives (472.5, 433.12, and 268.5 min at 50, 55, and $60^{\circ}C$, respectively) and the activation energy (9.85 kcal/mol) for the immobilized enzyme were higher than those for the free enzyme. Under the selected conditions, the immobilized enzyme had a higher $K_m$ (11.11 mM) and lower $V_{max}$ (105.26 U/mg protein) when compared with the free enzyme (8.33 mM and 125.0 U/mg protein, respectively). The operational stability of the biocatalyst was tested for both the hydrolysis of triglycerides and esterification of fatty acids with glycerol. After 4 cycles, the immobilized lipase retained approximately 50% and 80% of its original activity in the hydrolysis and esterification reactions, respectively.

Staphylococcal methicillin resistance expression under various growth conditions

  • Lee, Yoo-Nik;Ryoung, Poo-Ha;Lee, Young-Ik
    • Journal of Microbiology
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    • 제35권2호
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    • pp.103-108
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    • 1997
  • To improve the detection of methicillin resistant staphylococci, lowered incubation temperature (30.deg.) and inclusion of sodium chloride in media have been empirically recommended. However, in this study, we found that sodium chloride in Peptone-Yeast Extract-K$\_$2/HPO$\_$4/ (PYK) medium decreased methicillin minimum inhibitory concentrations. Divalent cations were shown to restore the expression of staphylococcal methicillin resistance. However, when it was determined by efficiency of plating, sodium chloride increased methicillin resistance expression on agar medium in which higher divalent cations were contained in the agar medium. The decrease of minimum inhibitory concentrations at 30.deg.C by sodium chloride occurred in Brain Heart Infusion but did not occur in other media investigated. Interestingly, both PYK and Brain Heart Infusion media had peptone, which contain cholic acids having detergent activities. Inclusion of sodium chloride in PYK caused a higher rate of autolysis. Penicillin binding protein 2a that has a low affinity to beta-lactam antibiotics, was highly inducible in methicillin resistant Staphylococcus epidermidis strains. In this study, we found that autolysins that are activated by the sodium chloride decreased the minimum inhibitory concentration at 30.deg.C, and peptidoglycan is weakened due to the presence of methicillin. Peptone in the media may aggravate the fragile cells. However, stabilization due to the presence of divalent cations and production of penicilin binding protein 2a increase the survival of staphylococci.

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전분폐수의 생물학적 처리에 관한 연구 1. 소맥 전분포수 처리균의 분리와 처리효과 (Biological Treatment of Starch Waste Part 1. Isolation of Wheat Starch Waste Decomposing Organisms and Their Efficiency on Waste Treatment)

  • 기우경
    • 한국미생물·생명공학회지
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    • 제3권3호
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    • pp.117-122
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    • 1975
  • Activated sluge를 폐수처리와 사료로 이용하기 위하여 고구마와 소맥 전분폐수에서 채집한 32종의 균원을 선별한 결과 2종의 우수한 균원을 확인하고 N-9균원으로부터 7종의 세균J-3으로부터 6종의 세균을 분리하여 최종 선별 결과 3종의 세균 J-3-1, J-3-2, J-9-2를 선별하였다. 1. N-9의 혼합 배양의 경우 J-3보다는 floc형성능이 좋았으며 N-9로부터 Eugelena속의 원생동물 1종이 J-3에서는 Bodo 속의 원생동물 1종이 동정되었다. 2. 3종의 선별된 균에 대한 형태학적 생리학적 특성을 검사했다. 3. Sample N-9의 혼합균 배양에 의해 99%의 폐수의 B.O.D가 제거되었으며 세균 단독 배양일 경우 92% 이상의 B.O.D.가 제거되었다. 4. 원료소맥 전분폐수의 B.O.D가 8,000 ppm이하에서도 B.B.D. 제거율과 sludge에서 생산된 protein 량은 폐수의 B.O.D. 와 처리하는 균에 따라 상이하였다.

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The Relationship between Prohibitin 1 Expression, Hepatotoxicity Induced by Acetaminophen, and Hepatoprotection by S-Adenosylmethionine in AML12 Cells

  • Eunhye Cho;Soohan Jung;Jina Kim;Kwang Suk Ko
    • Journal of Microbiology and Biotechnology
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    • 제32권11호
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    • pp.1447-1453
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    • 2022
  • Prohibitin 1 (Phb1) is a pleiotropic protein, located mainly in the mitochondrial inner membrane and involved in the regulation of cell proliferation and the stabilization of mitochondrial protein. Acetaminophen (APAP) is one of the most commonly used over-the-counter analgesics worldwide. However, at high dose, the accumulation of N-acetyl-p-benzoquinone imine (NAPQI) can lead to APAP-induced hepatotoxicity. In this study, we sought to understand the regulation of mRNA expression in relation to APAP and GSH metabolism by Phb1 in normal mouse AML12 hepatocytes. We used two different Phb1 silencing levels: high-efficiency (HE, >90%) and low-efficiency (LE, 50-60%). In addition, the siRNA-transfected cells were further pretreated with 0.5 mM of Sadenosylmethionine (SAMe) for 24 h before treatment with APAP at different doses (1-2 mM) for 24 h. The expression of APAP metabolism-related and antioxidant genes such as Cyp2e1 and Ugt1a1 were increased during SAMe pretreatment. Moreover, SAMe increased intracellular GSH concentration and it was maintained after APAP treatment. To sum up, Phb1 silencing and APAP treatment impaired the metabolism of APAP in hepatocytes, and SAMe exerted a protective effect against hepatotoxicity by upregulating antioxidant genes.

Molecular Cloning of a cDNA Encoding Putative Apolipophorin III from the Silkworm, Bombyx mori

  • Yun, Eun-Young;Goo, Tae-Won;Kim, Sung-Wan;Hwang, Jae-Sam;Kwon, O-Yu;Kang, Seok-Woo
    • 한국잠사학회:학술대회논문집
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    • 한국잠사학회 2003년도 International Symposium of Silkworm/Insect Biotechnology and Annual Meeting of Korea Society of Sericultural Science
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    • pp.85-86
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    • 2003
  • Apolipophorin III (apoLp-III) is a protypical exchangeable apolipoprotein that is abundant in hemolymph of many insect species. Its function lies in the stabilization of low-density lipophorin particles (LDLp) crossing the hemocoel in phases of high energy consumption to deliver lipids from the fat body to the flight muscle cells. But, recent studies with naive Galleria mellonella-apoLp-III gave first indications of an unexpected role of that protein in insect immune activation (Niere et al., 1999). (omitted)

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Effect of Cross-Linking Agents on L-Sorbose Production by Immobilized Gluconobacter suboxydans Cells

  • PARK, YOUNG-MIN;SANG-KI RHEE;EUI-SUNG CHOI;IN SIK CHUNG
    • Journal of Microbiology and Biotechnology
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    • 제8권6호
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    • pp.696-699
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    • 1998
  • Biological oxidation of D-sorbitol to L-sorbose using permeated and immobilized cells of Gluconobacter suboxydans was carried out to investigate the optimum reaction condition. The stabilization effect of cross-linking agents such as glutaraldehyde, tannic acid, and polyethylene imine to prevent the leakage of enzymes from beads containing permeated and immobilized cells of G. suboxydans was examined by the production of L-sorbose from the mixture of D-sorbitol and gluconic acid. The protein concentration effused from immobilized beads treated with only glutaraldehyde was $5.2\mug/m\ell$ after 20 h. The beads of G. suboxydans immobilized with alginate and cross-linked with 0.3% glutaraldehyde was the most useful for the oxidation of D-sorbitol to L-sorbose.

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Nerve growth factor-induced neurite outgrowth is potentiated by stabilization of TrkA receptors

  • Song, Eun-Joo;Yoo, Young-Sook
    • BMB Reports
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    • 제44권3호
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    • pp.182-186
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    • 2011
  • Exogenous stimuli such as nerve growth factor (NGF) exert their effects on neurite outgrowth via Trk neurotrophin receptors. TrkA receptors are known to be ubiquitinated via proteasome inhibition in the presence of NGF. However, the effect of proteasome inhibition on neurite outgrowth has not been studied extensively. To clarify these issues, we investigated signaling events in PC12 cells treated with NGF and the proteasome inhibitor MG132. We found that MG132 facilitated NGF-induced neurite outgrowth and potentiated the phosphorylation of the extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) and phosphatidylinositol-3-kinase (PI3K)/AKT pathways and TrkA receptors. MG132 stimulated internalization of surface TrkA receptor and stabilized intracellular TrkA receptor, and the $Ub^{K63}$ chain was found to be essential for stability. These results indicate that the ubiquitin-proteasome system potentiated neurite formation by regulating the stability of TrkA receptors.