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Showing papers on "Human serum albumin published in 1978"


Journal ArticleDOI
TL;DR: The carbodiimide-catalyzed conjugation of a 6700 molecular weight fragment of poly-L-lysine to radiolabeled human serum albumin or to horseradish peroxidase enhances the membrane transport of each protein into cultured mouse fibroblasts approximately 11- and 200-fold, respectively.
Abstract: The carbodiimide-catalyzed conjugation of a 6700 molecular weight fragment of poly-L-lysine to radiolabeled human serum albumin or to horseradish peroxidase enhances the membrane transport of each protein into cultured mouse fibroblasts approximately 11- and 200-fold, respectively. At least 50% of the peroxidase activity remained after conjugation. Trypsinization and carbamylation of the two conjugates demonstrates that the enhancement of their cellular uptake is related to their poly-L-lysine content. Simple addition to the medium of comparable amounts of free poly-L-lysine has no effect on the transport of either native protein. Addition of poly-L-ornithine (molecular weight 200,000) at 3-30 microgram/ml, a condition known to cause enhancement of 125I-labeled human serum albumin uptake by mouse sarcoma cells, has no visible effect on the cellular uptake of native horseradish peroxidase. The intracellular localization of the enzyme-poly-L-lysine conjugate can be demonstrated cytochemically by either light or transmission electron microscopy. A concentration of conjugate that increases the uptake more than 200-fold does not cause any detectable morphological change suggestive of cell toxicity. Furthermore, because poly-L-lysine is an excellent substrate for intracellular proteolytic enzymes, it can be expected to be broken down and reutilized in the cell.

212 citations


Journal ArticleDOI
TL;DR: High Performance Liquid Afinity Chromatography (HPLAC) and its Application to the Separation of Enzymes and Antigens

172 citations


Journal Article
TL;DR: For cardiac blood-pool imaging, Technetium-99m red blood cells labeled by an in vivo technique appears to be superior to the two Tc-HSA preparations studied.
Abstract: Technetium-99m red blood cells (Tc-RBC) labeled by an in vivo technique were compared with two preparations of Tc-99m human serum albumin (HSA) for cardiac blood-pool imaging. Relative distribution of the tracers was analyzed on end-diastolic frames of gated blood-pool studies and on whole-body (head to mid-thigh) anterior pinhole images. The Tc-RBC demonstrated greater relative percentage localization in the cardiac blood pool, higher target-to-background ratios in the left ventricle, and less liver concentration. For cardiac blood-pool imaging, Tc-RBC labeled by the in vivo approach appears to be superior to the two Tc-HSA preparations studied.

121 citations


Journal ArticleDOI
TL;DR: Urea (7M) and heating (35 to 55 degrees C) caused a major increase in the viscosity of mucin and mucin-albumin mixtures, suggesting that rupture of hydrogen bonds, unfolding and partial denaturation of mucIn promotes greater intertangling (possibly hydrophobic interactions) between mucIn and albumin molecules.
Abstract: The interaction of serum albumin with a model epithelial mucin from pig stomach was explored by rotary viscometry. During 30 min of incubation of human serum albumin(20mg/ml) and pig gastric mucin (8mg/ml) in iso-osmotic buffers at 37 degrees C, the solution became markedly viscous. Viscosity enhancement was proportional to albumin concentration (2-40mg/ml), was most pronounced under conditions of low shear rate (less than 45S-1), and was considerably greater than the additive or multiplicative viscosity values calculated from albumin or mucin solutions measured separately. The viscous mucin-albumin complex was destroyed by high shear rates (greater than 90S-1), but slowly re-formed under zero shear conditions. Elevation of pH (7 to 9), ionic strength (0.1 to 1.0), and addition of disodium EDTA (5mM) did not cause marked or specific alterations in the viscosity of the mixture, suggesting that electrostatic interactions probably do not stabilize mucin-albumin complexes. Urea (7M) and heating (35 to 55 degrees C) caused a major increase in the viscosity of mucin and mucin-albumin mixtures, suggesting that rupture of hydrogen bonds, unfolding and partial denaturation of mucin promotes greater intertangling (possibly hydrophobic interactions) between mucin and albumin molecules. The implications of mucin-albumin interaction in diseases associated with mucus obstruction are briefly discussed.

112 citations


Journal ArticleDOI
William T. Morgan1
TL;DR: It is of interest that copper, zinc, nickel, cadmium and cobalt effectively inhibit the binding of heme by alpha2-Histidine-rich glycoprotein, whereas other divalent metals tested, including calcium, magnesium and manganese do not appreciably affect the heme-alpha2-histidine- rich glycop protein interaction.

98 citations


Journal ArticleDOI
TL;DR: In this article, the binding to a biological macromolecule (human serum albumin, HSA) of small molecules (two drugs: warfarin and furosemide) has been studied by hig-performance liquid chromatography.

87 citations


Journal ArticleDOI
01 Oct 1978-Cancer
TL;DR: Lymphoid tissue of 42 patients with Hodgkin's disease was studied with immunohistological techniques and the presence of IgG in some Reed‐Sternberg (R‐S) cells was confirmed, and in addition serial sections and a double staining technique revealed that these cells also contained both kappa and lambda light chains.
Abstract: Lymphoid tissue of 42 patients with Hodgkin’s disease was studied with immunohistological techniques on the light microscopic and ultrastructural level. The presence of IgG in some Reed-Stemberg (R-S) cells was confirmed, and in addition serial sections and a double staining technique revealed that these cells also contained both kappa and lambda light chains. Furthermore two serum proteins, human serum albumin and alpha-1 -antitrypsin, were demonstrated in the same positive R-S cells. The ultrastructural localization of the immunoglobulin and of human serum albumin was not related to any protein synthesizing organelle or to structures related to endocytosis. It is suggested that the presence of immunoglobulin in R-S cells is the result of a disturbance of the cell wall integrity with subsequent nonspecific diffusion of immunoglobulin and other serum proteins into the cell. The presence of IgG therefore can not be taken as an argument for a B-cell origin of R-S cells. Possible mechanisms for the cell wall damage are discussed. Cancer 42:1793 -1803. 1978.

77 citations


Journal ArticleDOI
TL;DR: The seminal plasma in sperm suspensions from boar, bull, rabbit, ram and stallion was replaced with simple defined media as completely as possible by a combination of centrifugation through Ficoll and dilution, and sperm motility was preserved better in the presence of serum albumin.
Abstract: The seminal plasma in sperm suspensions from boar, bull, rabbit, ram and stallion was replaced with simple defined media as completely as possible by a combination of centrifugation through Ficoll and dilution. After this process, motility declined and the cells showed a tendency to agglutinate and/or stick to glass. Varying the ionic strength of the medium had little effect upon these parameters but sperm motility was preserved better in the presence of serum albumin. When a number of purified proteins and other macromolecules were tested individually in this way for their motility-preserving ability, bovine or human serum albumin was consistently the most effective. Defatting the albumin or altering its nature by mild reduction, oxidation or alkylation had little detectable effect on its motility-preserving ability; the protein did not appear to be acting as a chelator of metal ions, for it could not be replaced by EDTA. The response of the spermatozoa to replacemrnt of seminal plasma varied between species: bull spermatozoa were particularly sensitive and serum albumin had little effect upon their subsequent motility.

74 citations


Journal ArticleDOI
TL;DR: The results indicate that bilirubin is bound to lysine residue 240 at its high-affinity site on human serum albumin.
Abstract: Bilirubin can be coupled covalently to albumin by using water-soluble carbodi-imide as coupling reagent. The optimal specificity in the attachment of bilirubin to the high-affinity site on the albumin molecule was obtained by treating an albumin-bilirubin complex with carbodi-imide in low concentrations and for a short period. The product was reduced, carboxymethylated and digested with trypsin. By fractionation on Sephadex G-50 (superfine grade) a peptide fraction containing most of the bilirubin label was isolated. Further purification by paper chromatography gave one peptide, consisting of residues 240-258. The peptide containined a single lysine residue, 240, and had an intact disulphide bridge. The results indicate that bilirubin is bound to lysine residue 240 at its high-affinity site on human serum albumin.

74 citations


Journal ArticleDOI
TL;DR: The nuclear magnetic relaxation of water protons are used to measure the diffusional permeability (Pw) of human red blood cells to water as a function of concentration of nonpermeable and permeable solutes.
Abstract: We have used the nuclear magnetic relaxation of water protons to measure the diffusional permeability (Pw) of human red blood cells to water as a function of concentration of nonpermeable and permeable solutes. Measurements ofT1,T2, and a hybrid of the two were made and yielded the samePw. In the presence of the nonpermeable electrolyte NaCl, membrane permeability is constant between the volumes of 70 and 105μm3 and increases both as the cells swell and shrink beyond these limits. Changes in both the internal and external osmolarity, using the permeable solutes urea and ammonium chloride, do not affect membrane permeability. The composition of the suspending medium also has a significant effect on membrane permeability. Cells suspended in plasma have a cell water lifetime about 30% longer than cells of the same volume suspended in serum, or isotonic saline with human serum albumin. Addition of a crude preparation of fibrinogen in physiological amounts to isotonic saline and human serum albumin restores the cell water lifetime to a value similar to that observed in plasma.

68 citations


Journal ArticleDOI
TL;DR: The kinetics of bilirubin binding to human serum albumin at pH 7.40, 4 degrees C, was studied by monitoring changes in bilirubain absorbance and Curve-fitting procedures allowed the assignment of absorption coefficients to the intermediate species.

Journal ArticleDOI
TL;DR: Human serum albumin was labeled in various ways and with different radioactive labels and it appeared that all labeled HSA compounds showed a preferential adsorption onto PS (and SR) substrates.

Journal ArticleDOI
TL;DR: A derivative of the native-sequence tripeptide of the specific Cu(II)-transport site of human serum albumin, L-aspartyl-L-alanyl- L-histidine N-methylamide, was synthesized, and its binding to Cu( II) was examined to determine the influence of the side-chain groups on the Cu(ii) binding.
Abstract: A derivative of the native-sequence tripeptide of the specific Cu(II)-transport site of human serum albumin, L-aspartyl-L-alanyl-L-histidine N-methylamide, was synthesized, and its binding to Cu(II) was examined to determine the influence of the side-chain groups on the Cu(II) binding. The equilibria involved in the Cu(II)-L-aspartyl-L-alanyl-L-histidine N-methylamide system were investigated by analytical potentiometry. Three complex species were found in the pH range 4-10. The same species were identified in both the visible and circular-dichroism spectra. The main species present in the physiological pH range is shown to have the same ligands around the square-planar Cu(II) ion as those reported for albumin and tripeptides diglycyl-L-histidine and its N-methylamide derivative. The results obtained from competition experiments showed that this tripeptide has a higher affinity towards Cu(II) than has albumin itself. The overall findings are compared with those from albumin. At neutral pH the side chains do not play any important role in the Cu(II) binding, but at low pH the beta-carboxyl group of the N-terminal aspartic residue becomes important. A possible competition site on albumin for Cu(II) at low pH is discussed.


Journal ArticleDOI
TL;DR: Findings indicate the presence of a cooperative (allosteric) interaction between warfarin and the oleate ion for albumin binding and suggest a conformational transition in the albumin molecule as a result of interactions.

Journal ArticleDOI
TL;DR: The fractionation of crude hog pancreatic amylase by gradient elution was used to demonstrate the effect of alcohols as polarity-reducing agents and the properties required of materials used for hydrophobic interaction chromatography of biopolymers are discussed.



Journal ArticleDOI
TL;DR: A simple fluoroimmunoassay for the determination of albumin levels in serum, urine and cerebrospinal fluid employs magnetisable particles to which antibodies to human serum albumin are covalently linked, and albumin labelled with fluorescein.

Journal Article
TL;DR: The most accurate methods described to measure progesterone receptors in crude cytosol are those that use labeled progester one as ligand, provided that the nonspecific binding to corticosteroidbinding globulin is blocked by an excess of unlabeled cortisol.
Abstract: Progesterone receptors of human tumors of breast and gynecological origin were assayed with [6,7- 3 H]-17,21-dimethyl-19-norpregna-4,9-diene-3,20-dione or [1,2,6,7- 3 H]-progesterone plus a 100-fold excess of cortisol by the charcoal extraction method. Whereas both ligands gave concordant levels of binding for the high-capacity samples, higher nonspecific binding of 17,21-dimethyl-19-norpregna-4,9-diene-3,20-dione to serum albumin resulted in less precise estimates of low-binding-capacity samples. Ammonium sulfate precipitation of receptor and gradient ultracentrifugation analysis demonstrated that the nonspecific binding of 17,21-dimethyl-19-norpregna-4,9-diene-3,20-dione was nonprecipitable by 30% (NH 4 ) 2 SO 4 and migrated with serum albumin. That 17,21-dimethyl-19-norpregna-4,9-diene-3,20-dione binds to human serum albumin was further shown by gradient ultracentrifugation analysis and the charcoal extraction method. The importance of adding an excess of cortisol to block binding to corticosteroid-binding globulin with [1,2,6,7- 3 H]progesterone to measure progesterone receptor binding was redemonstrated. We conclude that presently the most accurate methods described to measure progesterone receptors in crude cytosol are those that use labeled progesterone as ligand, provided that the nonspecific binding to corticosteroidbinding globulin is blocked by an excess of unlabeled cortisol.

Journal ArticleDOI
TL;DR: The present study indicates that the fatty acid effect is chain length-dependent and competitively displace the binding of spin labels by serum albumin while fatty acids of longer chain length enhance the spin label binding allosterically.

Journal ArticleDOI
TL;DR: The findings suggest that a marked body protein depletion exists in chronic uremia, and that dietary treatment per se is not responsible for such a depleted state.

Journal ArticleDOI
TL;DR: It is shown that the rotational relaxation times of immunoglobulins exhibit a significant dependence on Immunoglobulin concentration, which is tentatively explained by loosening of the structure of the Fc part of the molecule in concentrations below 2 PM.

Journal ArticleDOI
TL;DR: Rabbit spermatozoa of freshly collected ejaculates were incubated in different fractions of seminal plasma proteins and motility was best maintained in the fractions which contained albumin, and rabbit serum albumin was more effective than bovine or human serumalbumin.
Abstract: Washed rabbit spermatozoa of freshly collected ejaculates were incubated in different fractions of seminal plasma proteins and examined for motility. Incubation media were protein fractions obtained by gel and ion-exchange chromatography. Motility was best maintained in the fractions which contained albumin, and rabbit serum albumin was more effective than bovine or human serum albumin. Concentrations of less than 4 mg rabbit serum albumin/ml resulted in markedly decreased motility.

Journal ArticleDOI
TL;DR: The apparent association constant of testosterone binding to pure human serum albumin was found to be significantly less at 5 g/dl (1.79 x 10(4) M-1), demonstrating that the common assumption that binding site characteristics are constant with serum protein dilution is not strictly valid.
Abstract: The apparent association constant of testosterone binding to pure human serum albumin was found to be significantly less at 5 g/dl (1.79 x 10(4) M-1) than at 1 g/dl (2.29 x 10(4) M-1). This demonstrates that the common assumption that binding site characteristics are constant with serum protein dilution is not strictly valid. Determination of the percent free testosterone at normal serum protein concentrations by equilibrium dialysis may also be in error by 20% or more due to fluid shifts within the test system.

Journal ArticleDOI
TL;DR: Binding to albumin may be more significant than binding to CBG in controlling the transfer rate of cortisol to the fetus in investigating in vitro in the perfused human placenta.
Abstract: The transfer and metabolism of cortisol and cortisone and the effect of protein binding on these processes have been investigated in vitro in the perfused human placenta. The clearance of cortisol in buffer, expressed as a fraction of the antipyrine transfer rate (clearance index), was 0.50 ± 0.05 SEM in either direction. Extensive conversion to cortisone (85%) occurred during transfer. Addition of corticosteroid-binding globulin (CBG) in amounts sufficient to bind 50% of the cortisol reduced the clearance (0.40 ± .026) insignificantly, whereas human serum albumin (HSA) in amounts sufficient to bind 50% of the cortisol reduced the clearance to 0.28 ± 0.012 (P <; 0.001) even though the association constant for albumin is approximately 1000-fold less. The percent of conversion to cortisone did not change significantly with protein binding. The clearance index of cortisone from a protein-free perfusate was 0.74. With CBG and albumin in the same concentrations as used in the cortisol experiments, the binding ...


Journal ArticleDOI
TL;DR: The reaction of the title compound with human serum albumin has been examined at various concentrations of the sulfonate andKinetic data suggest that there are two highly reactive lysine amino groups on the protein, five lysines which are less reactive and an undetermined number of additional nucleophilic groups that react very slowly with the reagent.

Journal ArticleDOI
TL;DR: Comparative studies of the T ELISA technique with bromocresol green, immunoturbidimetric and rocket immunoelectrophoretic methods were carried out and showed that TELISA could be used as an alternative method.

Journal ArticleDOI
TL;DR: The great difference of affinity of the serum albumins to parathion and paraoxon is discussed with respect to the fate ofParathion in the body.
Abstract: Binding of parathion and paraoxon to bovine serum albumin (BSA) and human serum albumin (HSA) was studied by using equilibrium dialysis. The concentration of unbound organophosphate was determined from its anticholinesterase activity. Binding of parathion to BSA was shown to be reversible. The organophosphates interact with only one type of binding sites in BSA and HSA. The affinity constants at pH 7.2 and 4° C for the interaction of BSA or HSA and parathion were found to be 2.7×106 and 1.5×106 M−1, respectively. The affinity constants for the interaction of the serum albumins and paraoxon were considerably lower, 6.0×103 and 1.6×104 M−1, respectively. Lowering the pH from 7.2 to 4.8 did not significantly affect the binding parameters. The great difference of affinity of the serum albumins to parathion and paraoxon is discussed with respect to the fate of parathion in the body.