• Title/Summary/Keyword: binding efficiency

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Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces

  • Renukaradhya K. Math;Nagakumar Bharatham;Palaksha K. Javaregowda;Han Dae Yun
    • Applied Microscopy
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    • v.51
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    • pp.17.1-17.10
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    • 2021
  • Our previous study on the binding activity between Cel5H and clay minerals showed highest binding efficiency among other cellulase enzymes cloned. Here, based on previous studies, we hypothesized that the positive amino acids on the surface of Cel5H protein may play an important role in binding to clay surfaces. To examine this, protein sequences of Bacillus licheniformis Cel5H (BlCel5H) and Paenibacillus polymyxa Cel5A (PpCel5A) were analyzed and then selected amino acids were mutated. These mutated proteins were investigated for binding activity and force measurement via atomic force microscopy (AFM). A total of seven amino acids which are only present in BlCel5H but not in PpCel5A were selected for mutational studies and the positive residues which are present in both were omitted. Of the seven selected surface lysine residues, only three mutants K196A(M2), K54A(M3) and K157T(M4) showed 12%, 7% and 8% less clay mineral binding ability, respectively compared with wild-type. The probable reason why other mutants did not show altered binding efficiency might be due to relative location of amino acids on the protein surface. Meanwhile, measurement of adhesion forces on mica sheets showed a well-defined maximum at 69±19 pN for wild-type, 58±19 pN for M2, 53±19 pN for M3, and 49±19 pN for M4 proteins. Hence, our results demonstrated that relative location of surface amino acids of Cel5H protein especially positive charged amino acids are important in the process of clay mineral-protein binding interaction through electrostatic exchange of charges.

Effects of Non-meat Protein Binders and Acidification on the Efficiency of Cold-Set Pork Restructuring by High Pressure

  • Hong, Geun-Pyo;Chun, Ji-Yeon;Lee, Si-Kyung;Choi, Mi-Jung
    • Food Science of Animal Resources
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    • v.32 no.3
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    • pp.301-307
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    • 2012
  • We investigated the effects of non-meat protein binders combined with glucono-${\delta}$-lactone (GdL) on the binding properties regarding restructured pork prepared by high-pressure treatment. Soy protein isolate (SPI), casein (CS), whey protein concentrate (WPC), and egg white (EW) were used as non-meat protein binders and compared with the control (no binder) and with the ${\kappa}$-carrageenan (KC) treatment. The compression and depression rates were 2.3 and 37 MPa/s, respectively, and pressurization was conducted at 200 MPa for 30 min at $4^{\circ}C$. After pressurization, the physical properties (pH, water-holding capacity, color, tensile strength, and microscopic structure) of the sample were evaluated. The combination of pressurization with acidification enabled cold-set meat binding, and the binding strength of restructured pork was enhanced by the addition of non-meat proteins. Among binders, SPI demonstrated the best efficiency in binding meat pieces. Therefore, the present study demonstrated that the combination of acidification and pressurization processes with the utilization of non-meat protein binders has a potential benefit in meat restructuring.

Authenticated Ticket-based Binding Update Protocol for Mobile IPv6 Network (모바일 IPv6 네트워크를 위한 티켓 기반의 인증된 바인딩 갱신 프로토콜)

  • Koo, Jung-Doo;Kim, Sang-Jin;Oh, Hee-Kuck
    • Journal of the Korea Institute of Information Security & Cryptology
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    • v.16 no.5
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    • pp.25-34
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    • 2006
  • Previous binding update protocols for Mobile IPv6 repeats the same protocol each time a mobile node moves to a foreign link Moreover, mobile nodes repeats the same protocol whenever the lifetime of the current binding update expires. To improve the efficiency of binding updates, we Propose a ticket-based binding update protocol for MIPv6. Our protocol minimizes the binding update cost using a ticket issued by the corresponding node. We have analyzed our protocol security against the security requirements of MIPv6 and existing attacks. Furthermore, we have also compared our protocol against previous binding update protocols.

Expression of an Angiogenin Binding Peptide and Its Anti-Angiogenic Activity

  • Choi, Suk-Jung;Ahn, Mi-Won;Yoon, Kyoung-Bum;Park, Jong-Won
    • BMB Reports
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    • v.31 no.5
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    • pp.427-431
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    • 1998
  • In the previous report (Choi et al., 1997), the angiogenin binding peptides identified from a phage-peptide library were analyzed by using the fusion proteins composed of the Escherichia coli maltose binding protein and its corresponding peptides. However, it was difficult to obtain a sufficient amount of the fusion proteins required for further analysis because of the low expression level. We now report a high level expression of the fusion protein and analysis of its anti-angiogenin activity. The use of strong T7 promoter and removal of signal sequence allowed about a 20-fold increase in the expression efficiency of the fusion protein. We were able to obtain about 10 mg of purified fusion protein from one liter of culture. The purified fusion protein showed angiogenin-specific affinity and inhibited the binding of biotinylated actin to human angiogenin at $IC_{50}$ of 0.6 mM. Its anti-angiogenin activity was also revealed by the chorioallantoic membrane assay.

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Influence of Alkali Metal Cation Type on Ionization Characteristics of Carbohydrates in ESI-MS

  • Choi, Sung-Seen;Kim, Jong-Chul
    • Bulletin of the Korean Chemical Society
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    • v.30 no.9
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    • pp.1996-2000
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    • 2009
  • Alkali metal salts were introduced to enhance the ionization efficiency of glucose and maltooligoses in electrospray ionization-mass spectrometry (ESI-MS). A mixture of the same moles of glucose, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose, and maltoheptaose was used. Salts of lithium, sodium, potassium, and cesium were employed as the cationizing agent. The ionization efficiency varied with the alkali metal cation types as well as the analyte sizes. Ion abundance distribution of the [M+$cation]^+$ ions of the carbohydrates varied with the fragmentor voltage. The maximum ion abundance at low fragmentor voltage was observed at maltose, while the maximum ion abundance at high fragmentor voltage shifted to maltotriose or maltotetraose for Na, K, and Cs. Variation of the ionization efficiency was explained with the hydrated cation size and the binding energy of the analyte and alkali metal cation.

Cloning and Iron Transportation of Nucleotide Binding Domain of Cryptosporidium andersoni ATP-Binding Cassette (CaABC) Gene

  • Wang, Ju-Hua;Xue, Xiu-Heng;Zhou, Jie;Fan, Cai-Yun;Xie, Qian-Qian;Wang, Pan
    • Parasites, Hosts and Diseases
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    • v.53 no.3
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    • pp.335-339
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    • 2015
  • Cryptosporidium andersoni ATP-binding cassette (CaABC) is an important membrane protein involved in substrate transport across the membrane. In this research, the nucleotide binding domain (NBD) of CaABC gene was amplified by PCR, and the eukaryotic expression vector of pEGFP-C1-CaNBD was reconstructed. Then, the recombinant plasmid of pEGFP-C1-CaNBD was transformed into the mouse intestinal epithelial cells (IECs) to study the iron transportation function of CaABC. The results indicated that NBD region of CaABC gene can significantly elevate the transport efficiency of $Ca^{2+}$, $Mg^{2+}$, $K^+$, and $HCO_3{^-}$ in IECs (P<0.05). The significance of this study is to find the ATPase inhibitors for NBD region of CaABC gene and to inhibit ATP binding and nutrient transport of CaABC transporter. Thus, C. andersoni will be killed by inhibition of nutrient uptake. This will open up a new way for treatment of cryptosporidiosis.

The Evaluation of Factors Which Influence Binding Efficiency of Modified in Vivo Erythrocyte Labeling Technique (변형 체내 표지법에 의한 적혈구 표지시 결합효율에 영향을 미치는 인자 평가)

  • Seo, Han-Kyung;Kim, Min-Woo;Lim, Seok-Tae;Sohn, Myung-Hee
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.4
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    • pp.300-305
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    • 2004
  • Purpose: We underwent this study to evaluate the factors which influence labeling efficiency when modified in vivo erythrocyte labeling technique was used. Materials and methods: Thirty healthy volunteers (M:F=19:11, age:$25{\pm}2$ yrs) were enrolled in this study. Totally, two hundred ten samples were obtained from them. The 1 mg of stannous pyrophosphate was injected intravenously at the beginning of labeling. After suitable tinning time (5 min, 20 min, 35 min) passed by, blood (5 mL, 3 mL or 1 mL) was withdrawn into 10 mL syringe previously containing Tc-99m (740 MBq) and anticoagulant (heparin, ACD or CPDA) through 19-gauged scalp needle. The generator ingrowth time of Tc-99m was within 24 hrs in each case. The blood samples were placed on rotating invertor during incubation (10 min, 25 min, 40 min) but some of them were not. Immediately after the conclusion of incubation, the labeled blood specimens to analyze were centrifuged. and then %Unbound Tc-99m was calculated. Statical analysis was used paired T-test and one way ANOVA with SPSS 10.0. Results: The binding efficiency at 1 mL of blood volume was $73{\pm}32%,\;91{\pm}10%$ at 3 mL and $96{\pm}7%$ at 5 mL (p<0.01). The binding efficiency at 5 min of tinning time was $45{\pm}23%,\;98{\pm}6%$, at 20 min and $97{\pm}8%$ at 35 min (p<0.001). The binding efficiency at 10 min of incubation time was $96{\pm}7%,\;95{\pm}12%$ at 25 min and $98{\pm}3%$ at 40 min (p>0.05). The binding efficiency in case of using rotating invertor was $96{\pm}7%$ and the binding efficiency in case of not using it was $87{\pm}18%$ (p>0.05). There was no significant difference between them. In binding efficiency according to kinds of anticoagulants, ACD was $98{\pm}4%$, CPDA was $97{\pm}6%$ and heparin was $89{\pm}20%$ (p<0.001). Conclusion: When modified in vivo erythrocyte labeling technique is used with Tc-99m, the methods to obtain the highest labeling efficiency are as follow. The withdrawing blood volume should be over 3 mL, tinning time should be kept between 20 min and 35 min, and incubation time should be kept between 10 min and 40 min. ACD or CPDA have to be used as a anticoagulant except heparin and the blood samples should be placed on rotating invertor during incubation.

The Search of Pig Pheromonal Odorants for Biostimulation Control System Technologies: A 2D-QSAR Model for Binding Affinity between 2-Cyclohexyloxytetrahydrofurane Analogues and Porcine Odorant Binding Protein (생물학적 자극 통제 수단으로 활용하기 위한 돼지 페로몬성 냄새 물질의 탐색: 2-Cyclohexyloxytetrahydrofurane 유도체와 Porcine Odorant Binding Protein 사이의 결합 친화력에 관한 2D-QSAR 모델)

  • Park, Chang-Sik;Choi, Yang-Seok;Sung, Nack-Do
    • Reproductive and Developmental Biology
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    • v.31 no.1
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    • pp.15-20
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    • 2007
  • To search of a new porcine pheromonal odorant for biostimulation control system technologies to offer a potentially useful and practical way to improve reproductive efficiency in livestock species, the two dimensional quantitative structure-activity relationship (QSAR) models between physicochemical parameters as descriptors of 2-cyclohexyloxytetrahydrofurane (A), 2-phenoxytetrahydrofurane (B) analogues and binding affinity constant ($p[Od.]_{50}$) for porcine odorant-binding protein (pOBP) as receptor of pig pheromones were derived and disscused. The statistical quality of the optimized 2D-QSAR model is good ($r^{2}=0.964$) and accounts for 96.4% of the variance in the binding affinity constants. It was found that the binding affinity constants were dependent upon the optimal value, $(SL)_{opt.}=1.418$ of substituent lipole (SL) in molecules. Therefore, the SL constant was very important factor for binding affinity.

Distributed Binding Update Scheme for Fast Handoff in Non-Nested Network Mobility Environment (비 중첩 네트워크 이동성 환경에서 빠른 핸드오프를 위한 분산 바인딩 갱신 기법)

  • Kim, Hoon;Song, Ui-Sung
    • The Journal of Korean Association of Computer Education
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    • v.10 no.5
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    • pp.81-88
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    • 2007
  • To support the movement of a mobile network consisting of several mobile nodes, the IETF NEMO working group proposed NEMO basic support protocol using bi-directional tunneling between the home agent and the mobile router. Since NEMO basic support protocol is based on bi-directional tunneling, all communications to and from mobile nodes are suffer from inefficient routing all the cases. To solve the routing inefficiency problem, several proposals are proposed. However, those proposals results in new problems, such as binding update storm and changes in existing protocol. Especially, binding update storm problem may cause the delay in binding update, this subsequently causes the difficulty for a real time service and the handoff delay on the movement of mobile network. In this paper, we propose a new binding update scheme which reduces binding update latency and the handoff delay through the distribution of the transmission time of binding update messages. The performance of the proposed scheme is also evaluated to demonstrate its efficiency. Simulation results show that the proposed scheme can efficiently reduce the binding update latency.

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Thermostable ${\beta}$-Glycosidase-CBD Fusion Protein for Biochemical Analysis of Cotton Scouring Efficiency

  • Ha, Jae-Seok;Lee, Young-Mi;Choi, Su-Lim;Song, Jae-Jun;Shin, Chul-Soo;Kim, Ju-Hea;Lee, Seung-Goo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.443-448
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    • 2008
  • Multidomain proteins for the biochemical analysis of the scouring efficiency of cotton fabrics were constructed by the fusion of a reporter moiety in the N-terminal and the cellulose binding domain (CBD) in the C-terminal. Based on the specific binding of the CBD of Cellulomonas fimi exoglucanase (Cex) to crystalline cellulose (Avicel), the reporter protein is guided to the cellulose fibers that are increasingly exposed as the scouring process proceeds. Among the tested reporter proteins, a thermostable ${\beta}$-glycosidase (BglA) from Thermus caldophilus was found to be most appropriate, showing a higher applicability and stability than GFP, DsRed2, or a tetrameric ${\beta}$-glycosidase (GUS) from Escherichia coli, which were precipitated more seriously during the expression and purification steps. When cotton fabrics with different scouring levels were treated with the BglA-CBD and incubated with X-Gal as the chromogenic substrate, an indigo color became visible within 2 h, and the color depth changed according to the conditions and extent of the scouring.