• BMB Reports
• /
• v.40 no.5
• /
• pp.723-730
• /
• 2007

Kinesin is an ATP-driven microtubule motor protein that plays important roles in control of microtubule dynamics, intracellular transport, cell division and signal transduction. The kinesin superfamily is composed of numerous members that are classified into 14 subfamilies. Animal kinesins have been well characterized. In contrast, plant kinesins have not yet to be characterized adequately. Here, a novel plant-specific kinesin gene, GhKCH2, has been cloned from cotton (Gossypium hirsutum) fibers and biochemically identified by prokaryotic expression, affinity purification, ATPase activity assay and microtubule-binding analysis. The putative motor domain of GhKCH2, $M_{396-734}$ corresponding to amino acids Q396-N734 was fused with 6$\times$His-tag, soluble-expressed in E. coli and affinity-purified in a large amount. The biochemical analysis demonstrated that the basal ATPase activity of $M_{396-734}$ is not activated by $Ca^{2+}$, but stimulated 30-fold max by microtubules. The enzymatic activation is microtubule-concentration-dependent, and the concentration of microtubules that corresponds to half-maximum activation was about 11 ${\mu}M$, much higher than that of other kinesins reported. The cosedimentation assay indicated that $M_{396-734}$ could bind to microtubules in vitro whenever the nucleotide AMP-PNP is present or absent. As a plant-specific microtubule-dependent kinesin with a lower microtubule-affinity and a nucleotide-independent microtubule-binding ability, cotton GhKCH2 might be involved in the function of microtubules during the deposition of cellulose microfibrils in fibers or the formation of cell wall.

• Asian-Australasian Journal of Animal Sciences
• /
• v.24 no.9
• /
• pp.1199-1203
• /
• 2011

The objective of this study was to examine the binding potential of sperm to the epithelium of the sperm storage tubules (SST) in vitro and in vivo to assess the functional maturation of fowl sperm. Sperm from the testis, epididymis, as well as the proximal, middle and distal vas deferens were incubated in vitro with either the uterovaginal junction (UVJ)- or infundibular tissue containing SST at $39^{\circ}C$ for 30 min. Aliquots of sperm were also artificially inseminated into the uteri of hens, and the UVJ and infundibulum were collected 24 h post artificial insemination (AI). After incubation and AI, tissues were washed to remove loosely adhered sperm and subjected to fluorescence staining with 4', 6-diamidino-2-phenylindole, dihydrochloride (DAPI) for counting the number of bound sperm per 0.25 mm2 of surface area. Sperm from the testis, epididymis, and the three segments of the vas deferens exhibited their differential (p<0.05) binding capacity, which increased gradually from the testicular to distal vas deferens sperm under both in vitro and in vivo conditions. Existing similar trend, sperm, regardless of their source had a lesser affinity to bind to the epithelium of the infundibular SST than to the UVJ-SST. These experimental results suggested that fowl sperm may undergo gradual changes in the process of functional maturation, whereby they gain the ability to bind to the epithelium of the SST during their passage through the male reproductive tract (MRT).

• 제어로봇시스템학회:학술대회논문집
• /
• 2005.06a
• /
• pp.1571-1576
• /
• 2005

DCS has many processing components and various communication elements. And its communication delay characteristic is affected diverse operating situation and context. Especially, binding signal which traversed from one control-node to another control-node undergo all sort of delay conditions. So its delay value has large deviation with the lapse of time, and the measurement of delay statistics during long time is very difficult by using general oscilloscope or other normal instruments. This thesis introduces the design and implementation of PC-based BDAS(Binding Delay Analysis System) System developed to overcomes these hardships. The system has signal-generator, IO-card, data-acquisition module, delay-calculation and analyzer module, those are implemented on industrial standard PC/Ethernet hardware and Windows/Linux platforms. This system can detect accurate whole-system-wide delay time including io, control processing and network delay, in the resolution of msec unit, and can analyze each channel's delay-historic data which is maintained by realtime database. So, this system has strong points of open system architecture, for example, user-friendly environment, low cost, high compatibility, simplicity of maintenance and high extension ability. Of all things, the measuring capability of long-time delay-statistics obtained through historic-DB make the system more valuable and useful, which function is essential to analyze accurate delay performance of DCS system. Using this system, the verification of delay performance of DCS for nuclear power plants is succeeded in KNICS(Korea Nuclear Instrumentation & Control System) projects

• BMB Reports
• /
• v.39 no.5
• /
• pp.560-570
• /
• 2006

Mistletoe (Viscum album) lectins, which are classified as a type II ribosome-inactivating protein (RIP) due to their unique biological function and the potential medical and therapeutic application in cancer cells, receive a rising attention. The heterodimeric glycoproteins contain the A-chain with catalytic activity and the B-chain with sugar binding properties. In recent years, studies involving the lectins from the white berry European mistletoe (Viscum album) and the yellow berry Korean mistletoe (Viscum album coloratum) have been described. However, the detailed mechanism in exerting unique cytotoxic effect on cancer cells still remains unclear. Here, we aim to understand and define the molecular basis and biological effects of the type II RIPs, through the studies of the recombinant Korean mistletoe lectin. To this end, we expressed, purified the recombinant Korean mistletoe lectin (rKML), and investigated its molecular characteristics in vitro, its cytotoxicity and ability to induce apoptotic cell death in cancer cells. To gain structural basis for its catalytic activity and sugar binding properties, we performed homology modeling studies based on the high degree of sequence identity and conserved secondary structure prediction between Korean and European, Himalayan mistletoe lectins, and Ricin.

• Journal of Ginseng Research
• /
• v.44 no.5
• /
• pp.690-696
• /
• 2020

Background: As the main metabolites of ginsenosides, 20(S, R)-protopanaxadiol [PPD(S, R)] and 20(S, R)-protopanaxatriol [PPT(S, R)] are the structural basis response to a series of pharmacological effects of their parent components. Although the estrogenicity of several ginsenosides has been confirmed, however, the underlying mechanisms of their estrogenic effects are still largely unclear. In this work, PPD(S, R) and PPT(S, R) were assessed for their ability to bind and activate human estrogen receptor α (hERα) by a combination of in vitro and in silico analysis. Methods: The recombinant hERα ligand-binding domain (hERα-LBD) was expressed in E. coli strain. The direct binding interactions of ginsenosides with hERα-LBD and their ERα agonistic potency were investigated by fluorescence polarization and reporter gene assays, respectively. Then, molecular dynamics simulations were carried out to simulate the binding modes between ginsenosides and hERα-LBD to reveal the structural basis for their agonist activities toward receptor. Results: Fluorescence polarization assay revealed that PPD(S, R) and PPT(S, R) could bind to hERα-LBD with moderate affinities. In the dual luciferase reporter assay using transiently transfected MCF-7 cells, PPD(S, R) and PPT(S, R) acted as agonists of hERα. Molecular docking results showed that these ginsenosides adopted an agonist conformation in the flexible hydrophobic ligand-binding pocket. The stereostructure of C-20 hydroxyl group and the presence of C-6 hydroxyl group exerted significant influence on the hydrogen bond network and steric hindrance, respectively. Conclusion: This work may provide insight into the chemical and pharmacological screening of novel therapeutic agents from ginsenosides.

• Bulletin of the Korean Chemical Society
• /
• v.27 no.11
• /
• pp.1801-1808
• /
• 2006

The interaction between whey carrier protein $\beta$-lactoglobulin type A and B (BLG-A and -B) and 2,2'-bipyridin n-hexyl dithiocarbamato Pd(II) nitrate (BPHDC-Pd(II)), a new heavy metal complex designed for anticancer property, was investigated by fluorescence spectroscopy combined with chemometry and circular dichroism (CD) techniques. A strong fluorescence quenching reaction of BPHDC-Pd(II) to BLG-A and -B was observed. Hence, BPHDC-Pd(II) complex can be bound to both BLG-A and -B, and quench the fluorescence spectra of the proteins. The quenching constant was determined using the modified Stern-Volmer equation. The binding parameters were evaluated by fluorescence quenching method. The results of binding study provided evidences presence of two and three sets of binding sites on the BLG-B and -A, respectively, for BPHDC-Pd(II) complex. Using fluorescence spectroscopy and chemometry, the ability of BLG-A and -B to form an intermediate upon interaction with BPHDC-Pd(II) complex was assessed. CD studies displayed that under influence of different concentrations of BPHDC-Pd(II) complex, the regular secondary structure of BLG-B had no significant changes, whereas for BLG-A a transition from $\alpha$-helix to $\beta$-structure was appeared. The results for both of BLG-A and -B displayed that BPHDC-Pd(II) complex can induce a conformational transition from the native form to an intermediate state with a slightly opened conformation, which is detectable with chemometry analyses.

• YAKHAK HOEJI
• /
• v.46 no.4
• /
• pp.219-225
• /
• 2002

We have performed a 3D-QSAR/CoMFA analysis of the cytotoxic activities of thirty-five 3-arylisoquinoline derivatives against SK-OV-3 tumor cell line. The results suggested that the electrostatic, steric and hydrophobic factors of 3-arylisoquinolines were strongly correlated with the antitumor activity. Considerable predictive ability (cross-validated r2 as high as 0.841) was obtained through CoMFA.

• Journal of Environmental Science International
• /
• v.7 no.1
• /
• pp.62-66
• /
• 1998

Toxic Mastoparan B(MP-B) which is purified from the venom of the hornet Vespa basalis is a cationic amphlphilic tetradecapeptide. MP-B and Its Ala-substituted analogues were synthesized by solld phase method and the toxic peptide-membrane interactions were examined by circular dichroism(CD) spectra, fluorescence spectra, and leakage abilities in phospholipid membranes. In the presence of phospholipid vesicles, synthetic MP-B and its analogues formed amphiphilic -helical structures, but in the buffer soletion, those exhibited random coil conformation as measured by CD. Fluorescence spectra of MP-B and its analogues which indicated the binding affinity of peptide on phospholipid vesicles showed that the replacement of Lys at position 2 and 11 with Ala caused a remarkable effect in the blue shalt and that at position 2, in the leakage ability of the peptide.

• Proceedings of the Korea Concrete Institute Conference
• /
• 2006.05b
• /
• pp.469-472
• /
• 2006

Amount used of admixture fare is increasing for concrete economic efficiency and ability security. Security of economic efficiency and ability can expect by use of this admixture fare but is displaying a lot of problems on early age strength hold by hydrate delay relatively. Specially, in the case of construction site, concrete strength can speak that interrelation is high with mold removal of forms time. Therefore, is economical and need examination of plan that can secure robber within 3 days using admixture fare such as fly ash and blast furnace slage differential speech to secure function. In this study, adding liquid admixture within 1% of a binding agent quantity to examine these problem with physical characteristic after hardening ago specially, strength change at standard and air dry curing of observed change in priority. Air dried and water curing total strength enhancement effects appeared by thing which is in case of add test result liquid admixture by below 1% and strength deputy by passage of age could know is not big.

• Proceedings of the KIEE Conference
• /
• 2003.07b
• /
• pp.777-779
• /
• 2003

This paper describes the real-time data linkage of the field control system for distributed control in nuclear power plant environment. The most important keys of digital control system in nuclear power plant are the reliability and stability of system, and real-time control ability. This Paper brought up the hardware construction using a new method about the design of each station located upon control transmission network to improve real-time ability of field control system, and measured the station binding time between devices connected to field control module. And it was confirmed performance improvement of overall system for real-time data linkage between control devices.