Spectrometric, Thermodynamic, pH Metric and Viscometric Studies on the Binding of TEALS as Surfactant with Albumin as Biopolymer

Background:: Since the interactions of small anions with protein are very important in their transportation and distribution processes in biological systems, it is helpful to study these interactions to understand the nature of the transportation and distribution processes. Therefore, it is aimed to study the interaction of albumin with surfactant molecule by different physical methods. Objective:: Present work attempts to work on assessing the structure, characterization of the surfactants as TEALS (tri-ethanalamine lauryl sulphate) binding sites, with albumin involved in various process of living being are discussed. Method:: The binding of surfactant TEALS to egg protein has been studied at different pH values and temperatures by spectrophotometric and equilibrium dialysis methods. The binding data were found to be pH and temperature dependent. The binding data studied by the absorbance method, were found approximately identical with those obtained from the equilibrium dialysis method. Results:: The association constants and the number of binding sites were calculated from Scatchard plots and found to be at maximum at lower pH and at lower temperature. The free energy of the combining sites was lowest at higher pH and highest at low pH. Therefore, a lower temperature and a lower pH offered more sites in the protein molecule for interaction with surfactant. The ΔG (free energies of aggregation) associated with the binding interaction of the surfactants and protein were calculated. The negative values of the ΔG confirm the feasibility of interaction between the surfactant and protein. All the observations recorded in this paper indicate that the TEALS has a good affinity of binding with egg protein and the number of binding sites is dependent on various physical and chemical factors. Conclusion:: On the basis of the results of the experiments which were conducted to examine the interaction between anionic surfactant and protein by measuring the various parameters of the solutions, it is concluded that the interaction of surfactant and protein gives an idea of fundamental understanding of the structure of surfactant-protein complex and their practical applications in every field.

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The Thermodynamic and Binding Studies of Hg+2 Ions with Egg Protein by Polarographic and pH Metric Techniques

Zeitschrift für Physikalische Chemie ◽

10.1515/zpch-2018-1158 ◽

2019 ◽

Vol 233 (8) ◽

pp. 1073-1090 ◽

Cited By ~ 2

Author(s):

Shveta Acharya ◽

Arun Kumar Sharma

Keyword(s):

Protein Molecule ◽

Binding Studies ◽

Temperature Dependent ◽

Ph Values ◽

Egg Protein ◽

Binding Data ◽

Number Of Binding Sites ◽

Lower Temperature ◽

Scatchard Plots ◽

Statistical Effects

AbstractThe binding of mercury (II) ion has been studied with egg protein at different pH values and temperatures by the polarographic technique. The binding data were found to be pH and temperature dependent. The intrinsic association constants (k) and the number of binding sites (n) were calculated from Scatchard plots and found tobe at the maximum at lower pH and at lower temperatures. The free energy change (ΔG°) of the combining sites were least at the higher pH and highest at the low pH; therefore, a lower temperature and lower pH offered more sites in the protein molecule for interaction with mercury (II) ions. Statistical effects seem to be more significant at lower mercury (II) ion concentrations, while at higher concentrations electrostatic effects and heterogeneity of sites are more significant.

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Abnormal Platelet Serotonin Uptake and Binding Sites in Myeloproliferative Disorders

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661099 ◽

1984 ◽

Vol 51 (03) ◽

pp. 349-353 ◽

Cited By ~ 9

Author(s):

C Caranobe ◽

P Sié ◽

F Fernandez ◽

J Pris ◽

S Moatti ◽

...

Keyword(s):

Binding Sites ◽

High Capacity ◽

Specific Inhibitor ◽

Myeloproliferative Disorders ◽

Normal Subjects ◽

Binding Data ◽

Full Explanation ◽

Number Of Binding Sites ◽

5 Ht Uptake ◽

Kinetics Of

SummaryA simultaneous investigation of the kinetics of serotonin (5 HT) uptake and of binding sites was carried out in the platelets of normal subjects and of 10 patients affected with various types of myeloproliferative disorders (MD). The 5 HT uptake was analysed according to the Lineweaver-Burk and the Eadie-Hofstee methods. With the two methods, the patient’s platelets exhibited a dramatic reduction of the Vi max and of the Km; in some patients the Eadie-Hofstee analysis revealed that a passive diffusion phenomenon is superimposed on the active 5 HT uptake at least for the higher concentration used. The binding data were analysed with the Scatchard method. Two classes of binding sites (high affinity - low capacity, low affinity - high capacity) were found in normal subjects and patients. Pharmacological studies with imipramine, a specific inhibitor of 5 HT uptake, suggested that both the sites are involved in 5 HT uptake. The number of both binding sites was significantly decreased in patient’s platelets while the affinity constants of these binding sites were not significantly reduced in comparison with those of the control subjects. No correlations were found between Vi max, Km and the number of binding sites. These results suggest that a reduction in the number of platelet membrane acceptors for 5 HT commonly occurs in myeloproliferative disorders but does not provide a full explanation of the uptake defect.

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THE PLATELET “REBOUND-PHENOMENON” DURING PROLONGED, CONTINUOUS PGI2 INFUSION OCCURS AT THE RECEPTOR LEVEL

10.1055/s-0038-1643452 ◽

1987 ◽

Author(s):

G Steurer ◽

H Sinzinger ◽

P Fitscha

Keyword(s):

Binding Sites ◽

Venous Blood ◽

Intermittent Infusion ◽

Receptor Level ◽

Binding Data ◽

Post Infusion ◽

Number Of Binding Sites ◽

Rebound Phenomenon ◽

Infusion Regimen

During earlier attempts in optimizing the therapeutic regimen with PGI2 we were able to discover an “ intra- and post-infusion platelet rebound” being characterized by an activated platelet function and a diminished responsiveness of platelets to the action of PGI2 in-vitro.In order to verify this phenomenon at the receptor level we infused continuously 6 patients suffering from peripheral vascular disease (PVD) with PGI2 at a rate of 5 ng/kg/min for 5 days. Anticoagulated venous blood has been drawn at different intervals. Saturation binding experiments on platelet membrane fraction have been performed using [3H]iloprost, a stable PGI2 analoque. Analysis of the binding data according to Scatchard demonstrated a decrease of receptor affinity with an increased number of binding sites.It is concluded, that intrainfusion rebound occurs at the receptor level, whereas the postinfusion rebound does not. This is a further piece of evidence that an intermittent infusion regimen is preferable.

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The Binding Of 125I-Von Willebrand Factor To Human Platelets In The Presence Of Ristocetin

10.1055/s-0038-1652985 ◽

1981 ◽

Author(s):

P Silber ◽

T H Finlay

Keyword(s):

Von Willebrand Factor ◽

Binding Sites ◽

Binding Constant ◽

Specific Binding ◽

Human Platelets ◽

Scatchard Analysis ◽

Binding Data ◽

Number Of Binding Sites ◽

Von Willebrand ◽

Willebrand Factor

The effect of ristocetin on the binding of 125I-porcine von Willebrand factor to human platelets was studied. Previously, we had shown that 125I-porcine von Willebrand factor binds to human platelets in the absence of ristocetin. The present work demonstrates that binding is stimulated by ristocetin and this stimulation is maximal at a ristocetin concentration of 2 mg/ml. At a ristocetin concentration of 0.5 mg/ml, Scatchard analysis indicates a binding constant of 5.18 × 10-9M and the presence of 105,000 binding sites. This compares with our previous finding, in the absence of ristocetin, of a binding constant of 2.92 × 10-7M and 4760 binding sites. These binding data assume the porcine von Willebrand factor to be a tetramer with a molecular weight of 9 × 105. This study indicates that ristocetin causes tighter binding and increases the number of binding sites on human platelets for porcine von Willebrand factor. Unlabelled porcine von Willebrand factor competitively inhibits the specific binding of the labelled protein and gives a binding constant of 0.17 × 10-9M. Similar results were obtained using human von Willebrand factor.

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Fluorescence-polarization studies on binding of 4-methylumbelliferyl β-d-galactopyranoside to Ricinus communis (castor-bean) agglutinin

Biochemical Journal ◽

10.1042/bj1910395 ◽

1980 ◽

Vol 191 (2) ◽

pp. 395-400 ◽

Cited By ~ 22

Author(s):

M I Khan ◽

M K Mathew ◽

P Balaram ◽

A Surolia

Keyword(s):

Binding Sites ◽

Fluorescence Polarization ◽

Castor Bean ◽

Ricinus Communis ◽

Equilibrium Dialysis ◽

Scatchard Analysis ◽

Polarization Data ◽

Appreciable Change ◽

Number Of Binding Sites ◽

Polarization Studies

The binding of Ricinus communis (castor-bean) agglutinin 1 to saccharides was studied by equilibrium dialysis and fluorescence polarization by using the fluorescently labelled sugar 4-methylumbelliferyl beta-D-galactopyranoside. No appreciable change in ligand fluorescence of 4-methylumbelliferyl beta-D-galactopyranoside was considerably polarized on its binding to the lectin. The association constants obtained by Scatchard analysis of equilibrium-dialysis and fluorescence-polarization data do not differ much from each other, and at 25 degrees C, Ka = 2.4 (+/- 0.2) X 10(4)M-1. These values agree reasonably well with that reported in the literature for Ricinus agglutinin 1. The number of binding sites obtained by the different experimental procedures is 1.94 +/- 0.1 per molecule of 120 000 daltons and is equal to the reported value of 2. The consistency in the values of Ka and number of binding sites indicate the absence of additional subsites on Ricinus agglutinin 1 for its specific sugars. In addition, the excellent agreement between the binding parameters obtained by equilibrium dialysis and fluorescence polarization indicate the potential of ligand-fluorescence-polarization measurements in the investigation of lectin-sugar interactions.

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Interfaces of Molecular Recognition Sites and Biosensors; Novel Bio-Sensing Materials

MRS Proceedings ◽

10.1557/opl.2015.575 ◽

2015 ◽

Vol 1793 ◽

pp. 1-6

Author(s):

Kyung M. Choi

Keyword(s):

Binding Sites ◽

Glass Surface ◽

High Sensitivity ◽

Affinity Binding ◽

Sensing Element ◽

Practical Applications ◽

High Affinity ◽

High Affinity Receptor ◽

Number Of Binding Sites ◽

Low Sensitivity

ABSTRACTWe introduce a sensing element, “Molecularly Imprinted Polymer (MIP),” which created by “Molecular Imprinted Technique.” However, the sensitivity of MIP’s based bio-sensors limits for practical applications due to the low sensitivity. To achieve a high sensitivity of MIP’s based sensors, the synthesis of “high affinity receptor or binding sites,” such as “monoclonal particles” is a key objective. In previous studies, affinity distribution plots indicated that “high affinity binding sites” were obtained when the number of binding sites per particle decreased. It means that smaller particles are expected to have higher affinity binding sites compared to larger particles. The result motivated us to produce small-sized MIP’s particles for the achievement of higher sensitivity. Microfluidic Synthesis has taken a great attention to synthesize small particles. However, the microfluidic synthesis gave us a difficulty, especially collections of MIP’s particles from the surface of PDMS-based microchannels due to a sticking problem. Thus, we employed a new approach, which can collect MIP’s particles without any sticking problem from the surface of the reactor. It is a photopatterned MIP’s system generated on the glass surface. We prepared a photomask with micro-sized patterns and then fabricate MIP’s particles on a glass surface by photopolymerization. Uniform MIP’s patterns were printed on the glass surface. The interface between the glass surface and the MIP’s pattern was observed by SEM. Micro-sized MIP’s particles were collected from the glass surface by scratching off the photocured MIP’s patterns.

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Equilibrium dialysis, ultrafiltration, and ultracentrifugation compared for determining the plasma-protein-binding characteristics of valproic acid.

Clinical Chemistry ◽

10.1093/clinchem/31.1.60 ◽

1985 ◽

Vol 31 (1) ◽

pp. 60-64 ◽

Cited By ~ 98

Author(s):

J Barré ◽

J M Chamouard ◽

G Houin ◽

J P Tillement

Keyword(s):

Valproic Acid ◽

Human Serum Albumin ◽

Binding Sites ◽

Plasma Proteins ◽

Equilibrium Dialysis ◽

Anticonvulsant Drug ◽

Drug Binding ◽

Binding Characteristics ◽

Affinity Constants ◽

Number Of Binding Sites

Abstract Equilibrium dialysis, ultrafiltration, and ultracentrifugation were compared to determine their reliability and applicability in the study of binding of an anticonvulsant drug, valproic acid, by plasma proteins. We studied drug binding with pooled serum and with solutions of human serum albumin at physiological concentrations. We compared binding characteristics such as number of binding sites, affinity constants, and percent of binding as measured by each method in the therapeutic range for valproic acid. Results by ultracentrifugation differed from those by equilibrium dialysis and ultrafiltration, which agreed reasonably well with each other.

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Binding of Ca++ to Human Prothrombin

10.1055/s-0039-1689427 ◽

1975 ◽

Author(s):

R. Benarous ◽

J. Elion

Keyword(s):

Binding Sites ◽

Equilibrium Dialysis ◽

Specific Property ◽

Binding Properties ◽

Binding Ability ◽

Ph Variation ◽

Maximum Value ◽

Number Of Binding Sites ◽

Scatchard Plots

The Ca++ binding properties of human prothrombin were studied by equilibrium dialysis using 45 calcium chloride at +4° C with prothrombin concentration of about 1 mg/ml equilibrated in 0.025 M Tris HCl, 0.12 M NaCl buffer pH 7.4. Scatchard plots obtained were similar to those described by Steenflo (1973) for bovine prothrombin, suggesting a positive cooperativity in the binding of Ca++ with a maximum ratio of bound Ca++/free Ca++ of 3 moles of Ca++ bound per mole of protein.The total number of binding sites was found to be at about 7, less than 10 to 12 found for bovine prothrombin. Ca++ binding was dependent on pH variation of the buffer with a maximum value for pH 8.5. Chemical modifications of carboxyl groups of prothrombin according to Hoare and Koshland (1967) abolished the Ca++ binding ability of the molecule confirming the essential role of these residues in this specific property of prothrombin.

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Abnormal Platelet Serotonin Uptake And Binding Sites In Myeloproliferative Disorders

10.1055/s-0038-1652398 ◽

1981 ◽

Author(s):

C Caranobe ◽

P Sié ◽

C Nouvel ◽

G Laurent ◽

J Pris ◽

...

Keyword(s):

Binding Sites ◽

High Capacity ◽

Myeloproliferative Disorders ◽

Binding Assays ◽

Dense Granules ◽

Plot Analysis ◽

Binding Data ◽

Plasmatic Membrane ◽

Number Of Binding Sites ◽

5 Ht Uptake

We previously shown that the platelets of patients suffering from myeloproliferative disorders (MD) present not only a reduced capacity to store serotonin (5 HT) in their dense granules but also a dramatic reduction in the initial velocity (Vi) of 5 HT uptake ; this suggests that the abnormalities are not restricted to the dense granules but involve the transport mechanism accross the platelet membrane.The present study concerns the 5 HT binding sites of MD platelets which present such a reduction of the Vi Max (Li- neweaver-Burke plot) of the 5 HT uptake. Binding assays were performed according to the method of Schik et al. (Biochem. Pharmacol. 1979, 28, 2667). Schatchard plot analysis of the binding data revealed two binding sites both in normals and patients : site A with a high affinity and a low capacity and site B with a low affinity and a high capacity.Thus the abnormal 5 HT transport accross the plasmatic membrane is the consequence of a quantitative reduction of the 5 HT binding sites and not of a qualitative defect of these sites. Nevertheless, in spite of the reduction of the number of binding sites, 5 HT-induced platelet aggregation was found normal in these patients.

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The Interaction and Thermodynamic Studies on the Binding of Congo Red Dye with Collagen Protein by Polarographic and Equilibrium Dialysis Techniques

Zeitschrift für Physikalische Chemie ◽

10.1515/zpch-2018-1181 ◽

2019 ◽

Vol 233 (5) ◽

pp. 691-705 ◽

Cited By ~ 6

Author(s):

Arun Kumar Sharma ◽

Shveta Acharya

Keyword(s):

Congo Red ◽

Equilibrium Dialysis ◽

Binding Studies ◽

Binding Interaction ◽

Effect Of Ph ◽

Congo Red Dye ◽

Collagen Protein ◽

Zinc Cadmium ◽

Number Of Binding Sites ◽

Red Dye

Abstract The survey of the existing literature revealed that the binding of Molybdenum, Vanadium, Zinc, Cadmium, Copper, Nickel and Cobalt with the protein is well known but no binding studies of Congo red molecules with collagen are reported. With a view to extend the existed knowledge of ecological nature of dye-protein system, it was thought of interest to investigate of properties of dye-protein mixture. The binding of Congo red dye has been studied with collagen protein using polarographic and equilibrium dialysis techniques. The intrinsic association constants and the number of binding sites have been calculated from Scatchard plots. The effect of pH on these constants was studied at pH 5.57, 7.50, 9.50 by polarographic technique and it was found that these values were decreased with increasing pH. The effect of pH was found to be similar by equilibrium dialysis technique. The values of different thermodynamic parameters have been reported. The free energies of aggregation, ΔG associated with the binding interaction of the dyes and protein were calculated. The negative values of the ΔG confirm the feasibility of interaction between the dye and protein.

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