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1.
The adsorption of protein films on polystyrene latex spheres was studied by optical mixing spectroscopy. With this technique, we show that both the hydrodynamic thickness of protein films and their optical density can be measured. Thus, we found that films of the glycoproteins isolated from the human erythrocyte membrane were four times as thick as films of either human serum albumin or bovine serum albumin for about the same surface coverage. This result suggests an end-on orientation for the adsorbed glycoprotein molecules, which is consistent with the model proposed by others for the orientation of these molecules at the surface of the red blood cell itself. 相似文献
2.
The binding of suramin to bovine and human serum albumin was investigated by gel filtration and spectroscopic measurements. Besides some low-affinity binding sites suramin has, on the bovine serum albumin molecule one and on the human serum albumin molecule two, high-affinity binding sites. Spectroscopic measurements reveal that there are large differences between the albumins in the mechanism of binding to the high-affinity binding sites. Further, it is suggested that high concentrations of suramin provoke an unfolding of the albumin moleculse. In order to explain the unusual behaviour of suramin in connection with the displacement of other ligands from the albumin binding the fluorescence probe 1-anilino-8-naphthalenesulfonic acid (ANS) was employed as a reporter group molecule for fluorescence as well as circular dichroism measurements. By these measurements it could be shown that suramin greatly influences the microorganization of both albumin molecules. In the case of these measurements large differences between bovine and human serum albumin were also found. 相似文献
3.
Thermodynamic parameters of the bovine serum albumin interaction with phosphatidyl choline liposomes have been determined. It is shown that the bovine serum albumin is adsorbed on the liposomal surface without change of the globule conformation, the polar interactions making dominant contribution in immobilization. The complexing is characterized by a binding constant and a great number of binding sites. 相似文献
4.
Iu P Tsarevski? B V Karal'nik 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》1975,(4):98-103
A study was made of the process of interaction between the tannin-treated sheep erythrocytes and the human serum albumin. Protein binding increased, whereas the loose/stable binding ratio decreased with the rise of the initial protein concentration. The process of stable albumin binding is described by the Langmour equation, this permitting to assess the limiting binding values -- about 6-10(5) molecules per erythrocyte. Albumin molecules were bound by erythrocyte with their greatest surface. At any concentration albumin was incapable of occupying more than 85% of the erythrocyte surface; at 90% binding level it blocked only 51--52% of the tannin-treated erythrocyte. The data obtained were regarded as a methodical basis for the preparation of erythrocytic diagnostic agents with the proteins of albumin type. 相似文献
5.
Total internal reflection/fluorescence photobleaching recovery study of serum albumin adsorption dynamics. 总被引:7,自引:5,他引:2
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The total internal reflection/fluorescence photobleaching recovery (TIR/FPR) technique (Thompson et al. 1981. Biophys. J. 33:435) is used to study adsorbed bovine serum albumin dynamics at a quartz glass/aqueous buffer interface. Adsorbed fluorescent labeled protein is bleached by a brief flash of the evanescent wave of a focused totally internally reflected laser beam. The rates of adsorption/desorption and surface diffusion determine the subsequent fluorescence recovery. The protein surface concentration is low enough to be proportional to the observed fluorescence and high enough to insure that the observed recovery rates arise mainly from adsorbed rather than bulk protein dynamics. The photobleaching recovery curves for rhodamine-labeled bovine serum albumin reveal both an irreversibly bound state and a multiplicity of reversibly bound states. The relative amount of reversible to irreversible adsorption increases with increasing bulk protein concentration. Since the adsorbed protein concentration appears to be too high to pack into a homogeneous surface monolayer, the wide range of desorption rates possibly results from multiple layers of protein on the surface. Comparison of the fluorescence recovery curves obtained with various focused laser beam widths suggests that some of the reversibly bound bovine serum albumin molecules can surface diffuse. Aside from their relevance to the surface chemistry of blood, these results demonstrate the feasibility of the TIR/FPR technique for measuring molecular dynamics on solid surfaces. 相似文献
6.
Different interactions of human and bovine serum albumin with Cibacron Blue and Blue Dextran 总被引:1,自引:0,他引:1
The interaction of human and bovine serum albumin with Cibacron Blue and Blue Dextran in aqueous solution was studied by means of difference spectroscopy. Both human and bovine albumin interact strongly with underivatized Cibacron Blue in three independent binding sites (K = 105). On the contrary, Blue Dextran interacts strongly only with human albumin, but does not bind appreciably to bovine albumin. These results suggest that the binding sites are exposed and easily accessible in human albumin, while in bovine albumin they are sterically hindered and therefore not accessible to the bulky Blue Dextran. 相似文献
7.
Hanh T. M. Phan Shannon Bartelt-Hunt Keith B. Rodenhausen Mathias Schubert Jason C. Bartz 《PloS one》2015,10(10)
Understanding protein adsorption kinetics to surfaces is of importance for various environmental and biomedical applications. Adsorption of bovine serum albumin to various self-assembled monolayer surfaces including neutral and charged hydrophilic and hydrophobic surfaces was investigated using in-situ combinatorial quartz crystal microbalance with dissipation and spectroscopic ellipsometry. Adsorption of bovine serum albumin varied as a function of surface properties, bovine serum albumin concentration and pH value. Charged surfaces exhibited a greater quantity of bovine serum albumin adsorption, a larger bovine serum albumin layer thickness, and increased density of bovine serum albumin protein compared to neutral surfaces at neutral pH value. The quantity of adsorbed bovine serum albumin protein increased with increasing bovine serum albumin concentration. After equilibrium sorption was reached at pH 7.0, desorption of bovine serum albumin occurred when pH was lowered to 2.0, which is below the isoelectric point of bovine serum albumin. Our data provide further evidence that combinatorial quartz crystal microbalance with dissipation and spectroscopic ellipsometry is a sensitive analytical tool to evaluate attachment and detachment of adsorbed proteins in systems with environmental implications. 相似文献
8.
The influence of bilirubin on fluidity and rotational correlation times of human erythrocyte membrane. 总被引:3,自引:0,他引:3
The effects of bilirubin on the membrane motion parameters of human erythrocyte membrane were determined by the spin labelled ESR method. It causes a decrease in the order parameter and an increase in the corresponding fluidity of the lipid molecules. Bovine serum albumin was found to inhibit effectively the effects due to bilirubin. The disturbance to the organization of membrane molecules by bilirubin as well as the protective effects of serum albumin are discussed on the basis of the experimental results. 相似文献
9.
The adsorption of proteins and protein--dodecyl sulphate complexes on N-(3-carboxypropionyl)aminodecyl-Sepharose.
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Equilibrium and kinetic aspects of the binding of several proteins to N-(3-carboxypropionyl)aminodecyl-Sepharose, an amphiphilic ampholytic adsorbent, were studied at 22 degrees C, pH 7.0, I 0.10--0.12. In the absence of detergents Scatchard plots are linear for human haemoglobin and soya-bean trypsin inhibitor, but non-linear for bovine serum albumin, which is also adsorbed more tightly than the other two proteins. The introducion of 3.5mM-sodium dodecyl sulphate causes dramatic increases in the amounts and affinities of serum albumin and haemoglobin adsorbed, but has relatively little effect on the trypsin inhibitor. At concentrations of sodium dodecyl sulphate greater than about 10mM there is a fall in the binding of all proteins, owing to competition from the detergent for binding sites on the adsorbent, and a tendency towards more uniform behaviour by different proteins. Kinetic experiments suggest that in the absence of the detergent haemoglobin and serum albumin are adsorbed initially by mainly ionic forces, but that subsequently hydrophobic forces become dominant. Addition of 3.5 mM-sodium dodecyl sulphate causes pronounced changes in the time course of adsorption of haemoglobin and serum albumin, the nature of the changes being different for each protein. The significance of these results is discussed. 相似文献
10.
Philip D. Morse 《Journal of cellular biochemistry》1974,2(1):60-70
The formation and stabilization of a complex between total erythrocyte apoprotein and monolayers of total erythrocyte lipid as measured by changes of surface pressure (Δπ) and rate of change of surface pressure (dπ/dt) was studied as a function of pH, ionic strength, and lipid surface pressure. Penetration of apoprotein into lipid monolayers was favored by conditions in which lipid and apoprotein were oppositely charged. Once the interaction was completed, the resultant surface complex was resistant to large changes in subphase pH and ionic strength as shown by the insensitivity of Δπ to these parameters. The dπ/dt, however, showed strong dependence on pH and ionic strength, but not on lipid surface pressure. A sharp decrease in dπ/dt around pH 3.5–4.5 is associated with the change in apoprotein charge from (+) to (?). Comparison of complex formation between apoprotein and bovine serum albumin, cytochrome c, and human hemoglobin suggests that erythrocyte apoprotein was specialized in its interaction with erythrocyte lipids. The data show that formation of an apoprotein-lipid complex at the air-water interface has both electrostatic and hydrophobic components. This contradicts results from other laboratories studying erythrocyte membrane recombination by bulk methods. 相似文献
11.
R A L?vstad 《The International journal of biochemistry》1986,18(2):171-173
Hemin-induced lysis of rat erythrocytes is markedly reduced by ceruloplasmin (human) and serum albumins from different species, the order of effectiveness beings: bovine albumin approximately equal to ceruloplasmin greater than human albumin approximately equal to dog albumin greater than apotransferrin (human). Although the proteins studied had hemin binding capacity, the best protective agents, ceruloplasmin and bovine albumin, did bind hemin less strongly than human and dog albumin. The results suggest the existence of another protective mechanism, possibly involving an interaction between erythrocyte membranes and serum proteins. 相似文献
12.
Adhesion of Listeria monocytogenes to silica surfaces after sequential and competitive adsorption of bovine serum albumin and beta-lactoglobulin.
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Adsorbed bovine serum albumin was resistant to exchange with beta-lactoglobulin, and when albumin was adsorbed from a mixture, its surface concentration increased with time. The passivating character of adsorbed albumin and its resistance to desorption were consistent with the level of Listeria monocytogenes adhesion evoked by albumin-containing protein films. 相似文献
13.
C K Luk 《Biopolymers》1971,10(7):1229-1242
The effect of cupric ion on the emission of tryptophan, tyrosine, and serum albumins is studied by emission spectroscopy and lifetime measurements. It is found that whenever cupric ion is bound to tryptophan or tyrosine, their emissions are quenched completely. The quenching may be due to an electron transfer mechanism. The fluorescence of complexes of cupric ions with serum albumins is partially quenched; this is because energy is transferred from tryptophan to the complexed cupric ions by a dipolar energy transfer mechanism. It is deduced from the present study that the tryptophan in the human serum albumin molecule is between 11 and 16 Å from the nearest eupric ion binding sites (assumed to be at the surface of the protein) and that one of the tryptophan in the bovine serum albumin molecule is very close to the cupric ion binding sites and the other is near the center of the bovine serum albumin molecule. It is also found that the deuterium solvent effect on serum albumin fluorescence is very small, and that the quenching of bovine serum albumin fluorescence at the N-F transition is the result of quenching of the fluorescence of both tryptophans. The phosphorescence lifetime apparatus, capable of measuring decay times of signals with intensities changing over a few orders of magnitude, and the ratio spectrofluorometer, both of which were constructed in this laboratory, are also described. 相似文献
14.
Circular dichroism studies on conformational changes in protein molecules upon adsorption on ultrafine polystyrene particles 总被引:1,自引:0,他引:1
The conformational changes in well-characterized model proteins [bovine ribonuclease A (RNase A), horseradish peroxidase, sperm-whole myoglobin, human hemoglobin, and bovine serum albumin (BSA)] upon adsorption on ultrafine polystyrene (PS) particles have been studied using circular dichroism (CD) spectroscopy. These proteins were chosen with special attention to molecular flexibility. The ultrafine PS particles were negatively charged and have average diameters of 20 or 30 nm. Utilization of these ultrafine PS particles makes it possible to apply the CD technique to determine the secondary structure of proteins adsorbed on the PS surface. Effects of protein properties and adsorption conditions on the extent of the changes in the secondary structure of protein molecules upon adsorption on ultrafine PS particles were studied. The CD spectrum changes upon adsorption were significant in the "soft" protein molecules (myoglobin, hemoglobin, and BSA), while they were insingnificant in the "rigid" proteins (RNase A and peroxidase). The soft proteins sustained a marked decrease in alpha-helix content upon adsorption. Moreover, the native alpha-helix content, which is given as the percentage of the alpha-helix content in the free proteins, of adsorbed BSA was found to decrease with decreasing pH and increase with increasing adsorbed amount. These observations confirm some well-known hypotheses for the confirmational chages in protein molecules upon adsorption. (c) 1992 John Wiley & Sons, Inc. 相似文献
15.
Reiter G Hassler N Weber V Falkenhagen D Fringeli UP 《Biochimica et biophysica acta》2004,1699(1-2):253-261
By in situ FTIR ATR measurements, the antibody (AB) recognition of human tumor necrosis factor-alpha (TNFalpha) immobilized on the Ge surface of a multiple internal reflection element (MIRE) was investigated. The experiments were performed in aqueous environment in a flow-through cell. After immobilization of TNFalpha on the Ge-MIRE by direct adsorption from aqueous solution, the immobilisate reached stability after about 1 h under flow-through conditions. The remaining sites of the Ge surface were saturated by bovine serum albumin (BSA) in order to prevent unspecific binding of anti-TNFalpha AB which was then added. The obtained FTIR ATR spectra were shown to result exclusively from AB specifically interacting with TNFalpha, since the absence of immunoglobulin binding to BSA adsorbed to the Ge MIRE was verified by a reference experiment. Finally, the stability of all adsorbed protein immobilisates was monitored under flow-through conditions for 10.5 h. The TNFalpha-AB complex showed a decrease of 7.4%, whereas the BSA adsorbate remained stable. IR measurements were performed with polarized light in order to study orientational effects of the immobilized proteins. The dichroic ratios and surface concentrations of all used proteins are available after quantitative analysis of the amide II bands. 相似文献
16.
Interactions between 1-benzoyl-4-p-chlorophenyl thiosemicarbazide and serum albumin: investigation by fluorescence spectroscopy 总被引:7,自引:0,他引:7
The interactions between 1-benzoyl-4-p-chlorphenyl thiosemicarbazide (BCPT) and bovine serum albumin (BSA) or human serum albumin (HSA) have been studied by fluorescence spectroscopy. By the analysis of fluorescence spectrum and fluorescence intensity, it was showed that BCPT has a strong ability to quench the intrinsic fluorescence of both bovine serum albumin and human serum albumin through a static quenching procedure. The binding constants of BCPT with BSA or HSA were determined at different temperatures based on the fluorescence quenching results. The binding sites were obtained and the binding force were suggested to be mainly hydrophobic. The effect of common ions on the binding constants was also investigated. A new fluorescence spectroscopy assay of the proteins is presented. The linear range is 5.36-67.0 microg mL(-1) with recovery of 101.1% for BSA, and the linear range is 8.28-144.9 microg mL(-1) with recovery of 102.6% for HSA. Determination of the proteins in bovine serum or in human serum by this method gives results which are very close to those obtained by using Coomassie Brilliant Blue G-250 colorimetry. A practical method was proposed for the determination of BCPT in human serum samples. 相似文献
17.
Polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) was applied to investigate the interaction of bovine serum albumin (BSA) and fibrinogen with a biomedical-grade 316LVM stainless steel surface, in terms of the adsorption thermodynamics and adsorption-induced secondary structure changes of the proteins. Highly negative apparent Gibbs energy of adsorption values revealed a spontaneous adsorption of both proteins onto the surface, accompanied by significant changes in their secondary structure. It was determined that, at saturated surface coverages, lateral interactions between the adsorbed BSA molecules induced rather extensive secondary structure changes. Fibrinogen's two coiled coils appeared to undergo negligible secondary structure changes upon adsorption of the protein, while large structural rearrangements of the protein's globular domains occurred upon adsorption. The secondary structure of adsorbed fibrinogen was not influenced by lateral interactions between the adsorbed fibrinogen molecules. PM-IRRAS was deemed to be viable for investigating protein adsorption and for obtaining information on adsorption-induced changes in their secondary structures. 相似文献
18.
H. Muzik M. E. Shea C. C. Lin H. Jamro S. Cassol L. M. Jerry L. Bryant 《In vitro cellular & developmental biology. Plant》1982,18(6):515-524
Summary Attempts were made to adapt human long-term B lymphoblastoid cell lines to prolonged growth in serum-free, chemically defined
media. A newly described medium, which is an enriched modification of Dulbecco’s modified Eagle’s medium containing additional
amino acids and vitamins, was used. The serum is totally replaced by albumin, transferrin, and soybean lipid. The cell lines
were all adaptable from RPMI 1640 over a period of time during which the 10% fetal bovine serum (FBS) concentration was reduced
and then eliminated in successive steps. After 3 to 6 wk minor alterations in cell shape and adhesion were noted without significant
histological changes. Growth characteristics were comparable in the new medium provided a double initial inoculum was used.
A panel of cell surface markers, including surface immunoglobulins, Ia antigens, Fc and complement receptors, and T and B
erythrocyte rosettes, all showed no altered expression. Molecular genotyping of Ia antigens was carried out by 2-D gel electrophoresis.
The antigens showed their full polymorphism without change and were shed into the new culture medium without alteration. Chromosome
analysis was performed on Q-banded karyotypes from one of the lines and showed no alteration resulting from the change to
serum-free conditions. Thus long-term B lymphoblastoid cell lines can be adapted to prolonged growth in serum-free medium.
This will facilitate the assay and isolation of cell products regulating lymphocyte function and the identification and characterization
of cell surface molecules free of interference from undefined serum components.
This work is supported by grants from the Medical Research Council of Canada, the National Cancer Institute of Cancer, and
the Alberta Heritage Fund. 相似文献
19.
用L-多聚赖氨酸、聚乙烯亚胺及L-多聚鸟氨酸三种多聚阳离子对壳聚糖进行共混修饰,制备了三种共混材料.在这些材料表面吸附了血清白蛋白,并利用圆二色(CD)光谱研究了白蛋白吸附到材料表面后的构象变化.结果显示,与天然状态相比,白蛋白吸附到共混材料表面后,其α-螺旋、β-折叠及无规则卷曲的含量均发生了明显改变.通过研究MC3T3-E1细胞在这些材料表面的生长情况,发现细胞的增殖与血清白蛋白的构象变化有一定关系,在吸附的白蛋白构象与天然构象最接近的共混材料表面,MC3T3-E1细胞增殖水平最高. 相似文献
20.
Iu P Tsarevski? B V Karal'nik 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》1975,(5):99-103
A study was made of a combined influence of the temperature of the medium and of sensitin concentration on the process of interaction of tannin-treated sheep erythrocytes and human serum albumin. On the basis of determination of the number of molecules bound by a single erythrocyte during hemosensitization,, depending on the mentioned conditions, it was revealed that the relative total albumin binding increased with the elevation of temperature. Elevation of temperature also led to the absolute and the relative increase in the stable and a reduction in the loose albumin binding by a single erythrocyte and the growth of sensitivity of the passive hemagglutination test. The role of chemical mechanisms in the erythrocyte albumin loading was demonstrated; this permitted to carry out erythrocyte albumin sensitization at a comparatively high temperature for the purpose of increasing the efficacy of passive hemagglutination test. 相似文献