• Title/Summary/Keyword: Molecular mechanism

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Characterization of Aspartate Aminotransferase Isoenzymes from Leaves of Lupinus albus L. cv Estoril

  • Martins, Maria Luisa Louro;De Freitas Barbosa, Miguel Pedro;De Varennes E Mendonca, Amarilis Paula Alberti
    • BMB Reports
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    • v.35 no.2
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    • pp.220-227
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    • 2002
  • Two aspartate aminoransferase (EC 2.6.1.1) isoenzymes (AAT-1 and AAT-2) from Lupinus albus L. cv Estoril were separated, purified, and characterized. The molecular weight, pI value, optimum pH, optimum temperature, and thermodynamic parameters for thermal inactivation of both isoenzymes were obtained. Studies of the kinetic mechanism, and the kinetics of product inhibition and high substrate concentration inhibition, were performed. The effect of some divalent ions and irreversible inhibitors on both AAT isoenzymes was also studied. Native PAGE showed a higher molecular weight for AAT-2 compared with AAT-1. AAT-1 appears to be more anionic than AAT-2, which was suggested by the anion exchange chromatography. SDS-PAGE showed a similar sub-unit molecular weight for both isoenzymes. The optimum pH (between 8,0 and 9.0) and temperature ($60-65^{\circ}C$) were similar for both isoenzymes. In the temperature range of $45-65^{\circ}C$, AAT-2 has higher thermostability than AAT-1. Both isoenzymes showed a high affinity for keto-acid substrates, as well as a higher affinity to aspartate than glutamate. Manganese ions induced an increase in both AAT isoenzymes activities, but no cooperative effect was detected. Among the inhibitors tested, hydroxylamine affected both isoenzymes activity by an irreversible inhibition mechanism.

Structure and Function of HtrA Family Proteins, the Key Players in Protein Quality Control

  • Kim, Dong-Young;Kim, Kyeong-Kyu
    • BMB Reports
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    • v.38 no.3
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    • pp.266-274
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    • 2005
  • High temperature requirement A (HtrA) and its homologues constitute the HtrA familiy proteins, a group of heat shock-induced serine proteases. Bacterial HtrA proteins perform crucial functions with regard to protein quality control in the periplasmic space, functioning as both molecular chaperones and proteases. In contrast to other bacterial quality control proteins, including ClpXP, ClpAP, and HslUV, HtrA proteins contain no regulatory components or ATP binding domains. Thus, they are commonly referred to as ATP-independent chaperone proteases. Whereas the function of ATP-dependent chaperone-proteases is regulated by ATP hydrolysis, HtrA exhibits a PDZ domain and a temperature-dependent switch mechanism, which effects the change in its function from molecular chaperone to protease. This mechanism is also related to substrate recognition and the fine control of its function. Structural and biochemical analyses of the three HtrA proteins, DegP, DegQ, and DegS, have provided us with clues as to the functional regulation of HtrA proteins, as well as their roles in protein quality control at atomic scales. The objective of this brief review is to discuss some of the recent studies which have been conducted regarding the structure and function of these HtrA proteins, and to compare their roles in the context of protein quality control.

DED Interaction of FADD and Caspase-8 in the Induction of Apoptotic Cell Death

  • Park, Young-Hoon;Han, Chang Woo;Jeong, Mi Suk;Jang, Se Bok
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.1034-1040
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    • 2022
  • Fas-associated death domain (FADD) is an adapter molecule that bridges the interaction between receptor-interacting protein 1 (RIP1) and aspartate-specific cysteine protease-8 (caspase-8). As the primary mediator of apoptotic cell death, caspase-8 has two N-terminal death-effector domains (DEDs) and it interacts with other proteins in the DED subfamily through several conserved residues. In the tumor necrosis receptor-1 (TNFR-1)-dependent signaling pathway, apoptosis is triggered by the caspase-8/FADD complex by stimulating receptor internalization. However, the molecular mechanism of complex formation by the DED proteins remains poorly understood. Here, we found that direct DED-DED interaction between FADD and caspase-8 and the structure-based mutations (Y8D/I128A, E12A/I128A, E12R/I128A, K39A/I128A, K39D/I128A, F122A/I128A, and L123A/I128A) of caspase-8 disrupted formation of the stable DED complex with FADD. Moreover, the monomeric crystal structure of the caspase-8 DEDs (F122A/I128A) was solved at 1.7 Å. This study will provide new insight into the interaction mechanism and structural characteristics between FADD and caspase-8 DED subfamily proteins.

Kinetics and Mechanism of Corrosion of ρ-alumina Bonded Alumina Castable by Molten Slag (ρ-알루미나결합 알루미나 캐스터블의 용융슬래그에 의한 침식기구)

  • 천승호;전병세
    • Journal of the Korean Ceramic Society
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    • v.40 no.10
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    • pp.1015-1020
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    • 2003
  • The Corrosion behavior of the matrix of the $\rho$-alumina bonded alumina vibrated castable was, on the basis of Jabsen's theory, elucidated by use of the Kingery's reaction mechanism. Corrosion of the matrix during induction period was controlled by the molecular diffusion. The temperature dependence of activation process is well established by the Arrhenius plots. The difference of Ca concentration between slag and interface is 23.2%, which causes a driving force of the materials transfer. The extent of the corrosion of the matrix is more deeper than that of the sintered mullite, but the corrosion mechanism can be well employed as the reaction mechanism proposed by the Kingery. The life time of the castable may be well estimated by the corrosion mechanism of Kingery.

Microscopic damping mechanism of micro-porous metal films

  • Du, Guangyu;Tan, Zhen;Li, Zhuolong;Liu, Kun;Lin, Zeng;Ba, Yaoshuai;Ba, Dechun
    • Current Applied Physics
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    • v.18 no.11
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    • pp.1388-1392
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    • 2018
  • Metal thin films are used widely to solve the vibration problem. However, damping mechanism is still not clear, which limits the further improvement of the damping properties for film and the development of multi-functional damping coating. In this paper, Damping microscopic mechanism of porous metal films was investigated at both macroscopically and microscopically mixed levels. Molecular dynamics simulation method was used to model and simulate the loading-unloading numerical experiment on the micro-pore and vacancy model to get the stress-strain curve and the microstructure diagram of different defects. And damping factor was calculated by the stress-strain curve. The results show that dislocations and new vacancies appear in the micro-pores when metal film is stretched. The energetic consumption from the motion of dislocation is the main reason for the damping properties of materials. Micro-mechanism of damping properties is discussed with the results of in-situ experiment.

Synthesis and Non-Isothermal Crystallization Behavior of Poly (ethylene-co-1,4-butylene terephthalate)s

  • Jinshu Yu;Deri Zhou;Weimin Chai;Lee, Byeongdu;Le, Seung-Woo;Jinhwan Yoon;Moonhor Ree
    • Macromolecular Research
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    • v.11 no.1
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    • pp.25-35
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    • 2003
  • A series of random poly(ethylene-co-1,4-butylene terephthalate)s (PEBTs), as well as poly(ethylene terephthalate) (PET) and poly(1,4-butylene terephthalate) (PBT), were synthesized by the bulk polycondensation. Their composition, molecular weight, and thermal properties were determined. All the copolymers are crystallizable, regardless of the compositions, which may originate from both even-atomic-numbered ethylene terephthalate and butylenes terephthalate units that undergo inherently crystallization. Non-isothermal crystallization exotherms were measured over the cooling rate of 2.5-20.0 K/min by calorimetry and then analyzed reasonably by the modified Avrami method rather than the Ozawa method. The results suggest that the primary crystallizations in the copolymers and the homopolymers follow a heterogeneous nucleation and spherulitic growth mechanism. However, when the cooling rate increases and the content of comonomer unit (ethylene glycol or 1,4-butylene glycol) increases, the crystallization behavior still becomes deviated slightly from the prediction of the modified Avrami analysis, which is due to the involvement of secondary crystallization and the formation of relatively low crystallinity. Overall, the crystallization rate is accelerated by increasing cooling rate but still depended on the composition. In addition, the activation energy in the non-isothermal crystallization was estimated.

Thimerosal generates superoxide anion by activating NADPH oxidase: a mechanism of thimerosal-induced calcium release

  • Kim, Eui-Kyung;Ryu, Sung-Ho;Suh, Pann-Ghill
    • Environmental Mutagens and Carcinogens
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    • v.22 no.4
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    • pp.229-235
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    • 2002
  • Thimerosal, a widely used preservative, has been well known to induce intracellular calcium mobilization in various cell types. However, the mechanism of its calcium mobilization is not clearly understood yet. For studying the mechanism of thimerosal-mediated calcium release, we have used HL60 cells in calcium-free Lockes solution that has no extracellular calcium. Thimerosal significantly reduced the lag period of initial calcium release whereas it enhanced the rate and magnitude of the calcium release in a dose-dependent manner. At the same time, we found that thimerosal generated superoxide anion by activating NADPH oxidase in dose- and time-dependent manner. Interestingly, the kinetics and the dosedependency of superoxide anion generation were very similar to those of intracellular calcium mobilization. In inhibitors study, the thimerosal-induced superoxide anion generation was significantly suppressed by DMSO as well as superoxide dismutase but not by genistein or EGTA. Surprisingly, the pretreatment with N-Acetyl-$_{L}$-Cysteine blocked almost completely the thimerosal-induced calcium increase, indicating that ROS playa key role in the calcium mobilization. The present results suggest that thimerosal-induced calcium mobilization is possibly mediated by the activation of NADPH oxidase and subsequent ROS generation.n.

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Mechanism of Methanol Conversion over Zeolite and Molecular Sieve Catalysts (제올라이트와 분자체 촉매에서 메탄올 전환 반응의 기구)

  • Seo, Gon;Min, Byung Goo
    • Korean Chemical Engineering Research
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    • v.44 no.4
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    • pp.329-339
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    • 2006
  • The production of lower olefins from methanol becomes an attractive process because of the rapid increase in crude oil price. This paper reivews the conversion mechanisms of methanol to hydrocarbons over zeolite and SAPO molecular sieve catalysts to understand the formation steps of lower olefins from methanol. The feasibility of the conversion mechanisms such as the direct mechanism based on well-defined intermediates and the hydrocarbon pool mechanism involving hydrocarbon moieties as an active centers is discussed with reepect to the induction period, the selectivity for products and the deactivation phenomena of the methanol conversion. The literature appeered since 1999 for the structure of the hydrocarbon pool and its catalytic role in the methanol conversion are summariged, and the prospect for the methanol-to-olefins process is described.

Functional Roles of Exosomes in Allergic Contact Dermatitis

  • Bocui Song;Qian Chen;Yuqi Li;Shuang Zhan;Rui Zhao;Xue Shen;Min Liu;Chunyu Tong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.12
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    • pp.1506-1514
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    • 2022
  • Allergic contact dermatitis (ACD) is an allergen-specific T-cell-mediated inflammatory response, albeit with unclear pathogenesis. Exosomes are nanoscale extracellular vesicles secreted by several cell types and widely distributed in various biological fluids. Exosomes affect the occurrence and development of ACD through immunoregulation among other ways. Nevertheless, the role of exosomes in ACD warrants further clarification. This review examines the progress of research into exosomes and their involvement in the pathogenesis, diagnosis, and treatment of ACD and provides ideas for exploring new diagnostic and treatment methods for this disease.