• Bulletin of the Korean Chemical Society
• /
• v.22 no.2
• /
• pp.145-148
• /
• 2001

The interaction between tetradecyl trimethyl ammonium bromide (TTAB) with bovine ${\alpha}-lactalbumin$ has been investigated at pH = 9 and at $37^{\circ}C$ by isothermal titration calorimetry, equilibrium dialysis and UV-Vis spectrophotometry methods. The binding data from unusual Scatchard plot have been analyzed in terms of the Hill equation for three sets of binding sites. The calorimetric data show that TTAB interacts endothermically with ${\alpha}-lactalbumin$ and causes protein unfolding below 2 mM concentration of TTAB, which is confirmed by spectrophotometric data. The unfolding of the protein would be mainly due to occupation of the second set of binding sites.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.36 no.6
• /
• pp.506-512
• /
• 1991

Monoclonal antibodies (mAb) to indole-3-acetic acid (IAA) were produced and characterized. Spleen cells from mouse immunized with IAA coupled to bovine serum albumin were fused with SP2/0-Ag14 myeloma cells. Three clones secreted specific antibodies to IAA were established to hybridoma cell lines and designated WLI-G1, WLI-G3 and WLI-Ell. The antibodies produced were classified into IgG, types and revealed the high degree of specificity by cross-reaction in the IAA derivatives and its analogues. In the IAA-ELISA with mAb, the measuring range of the assay was 1-500 p mol, and Ka and binding capacity calculated from Scatchard plot were 6.7 X 10$^{-10}$ L/M and 6 x 10$^{-10}$ L/M respectively. The ELISA with mAb can be used to quantitate IAA directly in crude plant eatract. The results showed that the immunoassay was easy and sensitive method to perform and applicate for quantitative analysis of IAA in plant.

• KSBB Journal
• /
• v.7 no.3
• /
• pp.201-208
• /
• 1992

Cell surface binding of epidermal growth factor(EGF) to EGF receptors was studied for a series of site-directed receptor mutants transfected into B82 mouse fibroblasts. Scatchard plots for truncation mutant receptors significantly lost nonlinearity for truncations below residue 1022. Transient plots of dissociation kinetics exhibited biphasic behavior for all receptor types, but the fraction of receptor in slow-dissociating form was reduced by an order of magnitude for the truncation mutants below residue 1022. Comparison of dissociation kinetics between control cells and cells treated with Triton X-100 revealed no significant variation for the slow-dissociating receptor form, but a noticeable variation was observed for the fast-dissociating receptor form when EGF receptors were truncated below residue 991. These results suggest that high affinity of EGF binding at cell surface depend on the EGF receptor cytoplasmic region.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1994.04a
• /
• pp.343-343
• /
• 1994

1. 목 적 : 혈액 중에 존재하는 약물은 대부분 혈장 단백질과 결합하며, 비단백 결합성 약물만이 생체막을 통과하여 여러 조직에 분포되고, target eel1에서 약리학적 작용을 나타내며, 대사, 배설 될 수 있다. 단백결합율이 높은 약물일수록 비결합성 약물의 양은 적어지며, 따라서 비결합성 약물의 증가는 약효의 상승을 의미하게 된다. 최근 만성 질환에 한약의 병용투여가 증가하고 있다. 본 실험에서는 단백결합율이 높은 감초의 주성분인 Glycyrrhizic acid(GA)와 고혈압 치료제로 많이 사용되는 칼슘 길항제를 병용 투여할 경우, 칼슘 길항제의 혈장 단백결합에 미치는 영향을 살펴 보았다. 2. 방 법 : Diltiazem hydrochloride, Verapamil hydrochloride, Nifedipine 와 GA를 model 약물로 하여 평형 투석법과 한외 여과법을 이용하여 fatty acid free human serum albumin(HSA), Low density lipoprotein( LDL ), of-Acid glycoprotein(AAG), plasma 각각에 대한 결합율을 HPLC로 분석하였으며 또한 Scatchard plot를 이용하여 binding parameter를 구하였다. 3. 결과 및 고찰 : GA는 Diltiazem의 HSA와 plasma의 결합율에 영향을 미쳤으며, Verapamil의 HSA, LDL, AAG, Plasma 결합율에, 그리고 Nifedipine의 HSA, LDL, Plasma의 단백 결합율에 영향을 주었으며, 각각 n과 Ka값에 변화를 주었다.

• Archives of Pharmacal Research
• /
• v.27 no.9
• /
• pp.978-983
• /
• 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.

• Archives of Pharmacal Research
• /
• v.14 no.3
• /
• pp.231-236
• /
• 1991

The high affinity binding sites for angiotensin II were solubilized from rat liver membranes by treatment with CHAPS. The binding protein was also partially purified by angiotensin III inhibitor-coupled Affi-gel affinity chromatography. Binding to the intact membrances as well as to the solubilized preparation was specific and saturable. According to the Scatchard plot, the membrane preparations exhibited a single class of high affinity binding sites with a Kd OF 0.71 nM. The solubilized preparation also showed the presence of a single class of bindings sites with less affinity (Kd of 14 nM). Meanwhile the competition studies using angiotensin II analogues represented two separate binding sites for angiotensin II and single binding site for antagonist. These latter findings were correlated to the results provided by Garrison's research group. More works are needed to clarify this discrepancy.

• Bulletin of the Korean Chemical Society
• /
• v.24 no.1
• /
• pp.45-48
• /
• 2003

A novel fluorimetric method has been developed for the determination of microgram quantities of bovine serum albumin (BSA) based on its enhancement effect of Nile Blue fluorescence at 670 nm, caused by binding of Nile Blue to BSA to produce a stable water soluble complex. The binding constant of micromole Nile Blue-BSA complex was estimated by Scatchard plot method. Under the optimal conditions, the increased fluorescence intensity was linearly related to BSA concentration in the range of 0.5-12.0 ㎍/mL. The detection limit was 0.2 ㎍/mL, and the relative standard deviation of six replicate measurements was 1.4% for 10.0 ㎍/mL BSA. There was little interference from amino acids, sugars and most of metal ions.

• Bulletin of the Korean Chemical Society
• /
• v.25 no.6
• /
• pp.791-795
• /
• 2004

The binding of cethyl trimethylammonium bromide, (CTAB) with human serum albumin (HSA) has been investigated at 5 mM phosphate buffer pH 7.0, 27 $^{\circ}C$ and various ionic strength using ion selective membrane electrodes. This method is faster and much more accurate than equilibrium dialysis technique, so provides sufficient and accurate data for binding data analysis. A novel and simple method was introduced for resolution and characterization of binding sets on basis of binding capacity concept. The values of Hill binding parameters were estimated for each set and used for calculation of intrinsic binding affinity. The results interpreted on basis of nature of forces which interfered in the interaction and represent the existence of three and two binding sets for binding of CTAB at $10^{-4}$ and $10^{-3}$ M of NaBr, respectively.

• Journal of the Korean Chemical Society
• /
• v.38 no.8
• /
• pp.585-589
• /
• 1994

The complexation of cadmium(II) with a well characterized local soil fulvic acid was investigated at different solution pH and fulvic acid concentration using ion selective electrode. The stability constants were determined using a continuous distribution model based on Scatchard Plot; this model takes into consideration the diversitv of cation binding sites on a naturally occurring organic macromolecules even if the nature of the binding sites may not be known. The mean value of $logKi(\mu)$ was found to increase at higher pH(pH 4.0: ${\mu}=3.79{\pm}0.74$ l\;mol^{-1}; pH 6.0: ${\mu}=4.51{\pm}0.78$ l\;mol^{-1}$), and in more dilute fulvic acid concentration([FA] = 50 mgl$^{-1}: {\mu}=4.16{\pm}0.60$ l\;mol^{-1}; [FA]=151 mgl$^{-1}:{\mu}=3.75{\pm}0.5\;l\;mol^{-1}$). The intrinsic constant(logKint) for binding at the strongest site was measured to be around $4.72\;l\;mol^{-1}$ at both pH4.0 and 5.0, but was found to be increased to $6.03\;l\;mol^{-1}$ when pH was 6.0.

• Bulletin of the Korean Chemical Society
• /
• v.15 no.7
• /
• pp.581-584
• /
• 1994

A comparative investigation of the complexations of divalent metal ions (Cd, Cu, and Pb) by a well characterized soil humic acid (HA) from Okchun Metamorphic Belt was carried out in 0.05 M $KNO_3$ and pH of 4.5 using ion selective electrodes. A continuous distribution model based on the Scatchard Plot was used to determine the stability constants, because the constants obtained by this modeling technique takes the variations in binding energies into consideration without regards to the manner in which M(II) ion is bound on HA. The mean value of log $K_i$ were $4.05{\pm}0.60,\;4.92{\pm}0.36,\;and\;5.63{\pm}0.34\;{\ell}\;mol^{-1}$ for Cd(II)-, Pb(II), and Cu(II)-humate complexes respectively. The values of intrinsic constant (log $K_{int}$; binding at strongest site) were $7.12{\pm}0.30,\;6.59{\pm}0.32,\;and5.07{\pm}0.56\;{\ell}\; mol^{-1}$ in the order Cu(II) > Pb(II) > Cd(II) ion.