• Journal of Pharmaceutical Investigation
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• 제23권3호
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• pp.119-125
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• 1993

The protein binding of salicylate analogs has been investigated by equilibrium dialysis. A series of binding experiments were performed in order to elucidate the effects of physicochemical properties of salicylate analogs on the binding with bovine serum albumin. Attempts to correlate affinity constants with capacity factor, steric factor and Hammett ${\sigma}$ values suggested hydrophobic forces to be involved in the binding of salicylate analogs. Steric factor contributes to binding process partly, whereas electronic interaction appears to be insignificant.

• KSBB Journal
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• 제13권2호
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• pp.125-132
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• 1998

Protein affinity membrane was prepared via the coating of chitosan gel on the porous flat polysulfone membrane surface, followed by the immobilization f the reactive dye (Cibacron Blue 3GA) to the chitonsan gel. The maximum protein binding capacity of affinity membrane was about 70${\mu}g/cm^2$ determined by the batch adsorption experiments of human serum albumin (HSA). Using module of this membrane, the characteristics of protein chromatography were investigated through the experiments of elution and frontal chromatography of HSA. This membrane module promises as a chromatography column, since it represented a lower pressure drop and a greater reproducibility. The protein separation ratio was significantly influenced by the flow rate of mobile phase and the injection quantity of HSA. The dynamic protein binding capacity of module decreased from the equilibrium binding capacity with increasing flow rate and approached the value of 15 - 20 ${\mu}g/cm^2$ for flow rates above 6 mL/min.

• Archives of Pharmacal Research
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• 제27권9호
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• pp.978-983
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• 2004

The effects of glycyrrhizic acid (GLZ) on protein binding of diltiazem, verapamil, and nifedipine were investigated. Protein binding studies (human serum, human serum albumin (HSA) and (X1-acid glycoprotein (AAG)) were conducted using the equilibrium dialysis method with and without addition of GLZ. The binding parameters, such as the number of moles of bound drug per mole of protein, the number of binding sites per protein molecule, and the association con-stant, were estimated using the Scatchard plot. The serum binding of nifedipine, verapamil, and diltiazem was displaced with addition of GLZ, and the decreases of Ks for serum were observed. GLZ decreased the association constants of three drugs for HSA and AAG, while the binding capacity remained similar with addition of GLZ. Although the characteristics of interaction were not clear, GLZ seemed to mainly affect HSA binding of nifedipine rather than AAG binding, while GLZ seemed to affect both AAG- and HSA-bindings of verapamil and dilt-iazem resulting in a serum binding displacement.

• Preventive Nutrition and Food Science
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• 제15권4호
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• pp.282-286
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• 2010

To isolate a calcium-binding peptide from chlorella protein hydrolysates, chlorella protein was extracted and hydrolyzed using Flavourzyme, a commercial protease. The degree of hydrolysis and calcium-binding capacity were determined using trinitrobenzenesulfonic acid and orthophenanthroline methods, respectively. The enzymatic hydrolysis of chlorella protein for 6 hr was sufficient for the preparation of chlorella protein hydrolysates. The hydrolysates of chlorella protein were then ultra-filtered under 5 kDa as molecular weight. The membrane-filtered solution was fractionated using ion exchange, reverse phase, normal phase chromatography, and fast protein liquid chromatography to identify a calcium-binding peptide. The purified calcium-binding peptide had a calcium binding activity of 0.166 mM and was determined to be 700.48 Da as molecular weight, and partially identified as a peptide containing Asn-Ser-Gly-Cys based on liquid chromatography/electrospray ionization tandem mass spectrum.

• Journal of Nutrition and Health
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• 제32권6호
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• pp.628-636
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• 1999

There is a marked increase in geriatric disease, especially liver disease, due to the continuous increase in alcohol and fat consumption. Since the fatty liver, induced by alcohol or fat, is basically from abnormalities in the lipid metabolism, it is possible that fatty acid binding protein(FABP) which is related to the fatty acid metabolism may also be abnormal in these livers. FABP is a small molecular weight protein family present in cytosol in high concentration. It has been proposed as a fatty acid transfer protein and as a binding protein responsible for controlling intracellular free fatty acid concentration. In this research, we have examined the relationship between liver FABP and fatty liver induced by alcohol or high cholesterol diet. Rats were fed one of either semipurified liquid diets; control diet containing 65% carbohydrate, 20% protein, and 15% fat or high cholesterol diet containing 1%(w/w) cholesterol or alcohol diet containing 37% of alcohol instead of carbohydrate. After 5 weeks of feeding period, all rats received commercial chow diet for 5 weeks to examine recovery effect. Liver and blood samples were collected at 0, 1, 3, 5 and 10 weeks to analyze lipid compositions. FABP was purified from liver cytosol and injected to rabbit to obtain antiserum. Liver FABP amount was determined by SDS-PAGE and western blotting methods. Fatty acid binding capacity was determined by binding of 14Cpalmitate with the delipidated liver cytosol. Consumption of alcohol increased serum cholesterol, triglyceride concentration and decreased HDL-cholesterol concentration after 5 weeks. Serum apolipoprotein B concentration increased after 3 weeks and LDL-cholesterol and apolipoprotein A concentration changed after 1 week. Liver cholesterol and triglyceride concentration increased after 3 weeks. Consumption of high cholesterol diet changed liver and serum lipid composition after 3 weeks. Swiching to normal diet for 5 weeks did not normalize most of lipid composition in serum and liver except serum and liver except serum cholesterol, triglyceride and liver cholesterol. Liver cytosol FABP content and the fatty acid binding capacity decreased dramatically after 1 week with alcohol consumption. This results indicate that FABP content changes before the changes before the changes of blood or liver lipid composition, suggesting changes of FABP may cause development of the fatty liver induced by alcohol and can be used as an index of detecting a early development of fatty liver.

• 한국축산식품학회지
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• 제32권3호
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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.

• BMB Reports
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• 제33권5호
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• pp.370-378
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• 2000

Incubation of purified laminin1-nidogen1 complexes with $[{\gamma}-^{32}P]-ATP$ in the presence of the catalytic subunit of the protein kinase A (cAMP-dependent protein kinase) resulted in the phosphorylation of the alpha chain of laminin-1 and of the nidogen-1 molecule. Aminoacid electrophoresis indicated that phosphate was incorporated on serine residues. The phosphorylation effect of laminin-1 on the process of self assembly was studied by turbidometry. In these experiments, the phosphorylated laminin-1 showed a reduced maximal aggregation capacity in comparison to the non-phosphorylated molecule. Examination of the laminin-1 network under the electron microscope showed that the phosphorylated sample formed mainly linear extended oligomers, in contrast to controls that formed large and dense multimeric aggregates. Heparin binding on phosphorylated laminin-1 in comparison to controls was also tested using solid-phase binding assays. The results indicated an enhanced heparin binding to the phosphorylated protein. The results of this study indicate that laminin1-nidogen1 is a substrate for protein kinase A in vitro. This phosphorylation had an obvious influence on the lamininl-nidogen1 network formation and the heparin binding capacity of this molecule. However, further studies are needed to investigate whether or not this phenomenon could play a role in the formation of the structure of basement membranes in vivo.

• 멤브레인
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• 제19권2호
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• pp.113-121
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• 2009

실리카 입자를 기공 형성제로 사용하여 물리적 강도와 단백질 결합용량이 높은 다공성 키토산 및 키틴 친화 막을 제조하였다. 키토산 친화 막의 BSA 단백질 결합용량은 최대 21.8mg/mL이었으며, 키틴 친화 막의 lysozyme 효소 결합용량은 최대 26.1mg/mL이었다. 제조된 다공성 키토산 및 키틴 친화 막을 사용하여 단백질 용액의 loading 유량, loading 양 및 농도 변화에 따른 BSA와 lysozyme의 친화 막 여과 크로마토그래피 분리 실험을 수행하였다. 친화 막 여과 크로마토그래피 분리 실험을 통해 얻어진 loading/washing/elution의 단계로 구성된 일련의 크로마토그램으로부터 단백질 용출량과 결합수율을 구하였다. 키토산 및 키틴 친화 막에의 BSA 및 lysozyme 단백질의 결합량과 결합수율은 loading용액의 유량이 작을수록, 주입량 및 농도가 클수록 증가하였다. 이 결과로부터 실리카 입자를 기공 형성제로 사용하여 제조된 다공성 키토산 및 키틴 막은 단백질의 대규모 여과 크로마토그래피 분리를 위한 친화 막으로서 효과적인 활용이 기대된다.

• 한국식품영양학회지
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• 제7권2호
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• pp.128-136
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• 1994

Mackerel muscle protein hydrolysates, which were prepared from defatted mackerel meal by proteases such as complex enzyme, alcalase, bromelain, pancrease, pepsin, w-chymotrypsin, trypsin and papain, were tested for the antioxidative action against linoleic acid. Among proteases tested, the hydrolysates obtained from the treatment of complex enzyme, bromelain and alcalase showed higher antioxidative effects. Also, the hydrolysates showed the synergistic effects with o-tocopherol and the inhibitory effects for peroxidation of metal ions(Fe3+, Cua+) From the profiles of fractionation of the hydrolysates with Bio-gel P-2 column, the most active fractions, part I(complex enzyme-derived) and part e(bromelain-derived), had below MW 1,400 and the antioxidative effects were closely related to the binding capacity with metal ion(Cua+). Amno acid composition of the part I was abundant in histidine, arginine, phenylalanine and lysine, and the part e was abundant in lysine, glutamic acid and leucine.

• Journal of Microbiology
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• 제33권3호
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• pp.228-233
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• 1995

As S. epidermidis cell was fractionated into cell wall, cell membrane, and cytoplasm, the cell membrane proved to be the most efficient absorbent for lead ion. Utrasonication was effective, when the cells were treated during their exponential growth. The amount of the lead ion adsorbed in cell membrane decreased as hydrogen ion concentration of solution increased. Protein purified from the cell membrane showed higher adsorption capacity for the lead ion than peptidoglycan, teichoic acid from cell wall, or cell membrane lipid. Modification of carboxyl groups in the membrane protein with ethylenediamine and 1-ethyl-3-carbodiimide hydrochloride resulted in a considerable decrease of lead ion adsorption capability, suggesting that the main binding site for lead ion was the carboxyl groups of protein in cell membrane.