Gel chromatographic procedure that corrects for Donnan effects in studies of ligand binding. Its application to the interaction of copper ions with bovine serum albumin and deoxyribonucleic acid

Biochemistry ◽  
1974 ◽  
Vol 13 (9) ◽  
pp. 1832-1836 ◽  
Author(s):  
D. O. Jordan ◽  
S. J. Lovell ◽  
D. R. Phillips ◽  
D. J. Winzor
Keyword(s):  
Bovine Serum Albumin ◽  
Ligand Binding ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Deoxyribonucleic Acid ◽  
Copper Ions ◽  
Chromatographic Procedure

Über Desoxyribonucleinsäure-Molekeln in Protein-Mischfilme

1959 ◽  
Vol 14 (12) ◽  
pp. 770-779 ◽  
Author(s):  
A. Kleinschmidt ◽  
H. Rüter ◽  
W. Hellmann ◽  
R. K. Zahn ◽  
A. Docter ◽  
...  
Keyword(s):  
Electron Microscope ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Deoxyribonucleic Acid ◽  
Carbon Support ◽  
Dna Molecules ◽  
Langmuir Trough ◽  
Surface Monolayers

A nucleoprotein of a vitrous consistency was extracted from the gonads of the coalfish (Gadus virens).The preparation of deoxyribonucleic acid (DNA) from this nucleoprotein and from staphylococci is described. Both of these different kinds of DNA have been mixed with bovine serum albumin or cytochrom c respectively to produce solutions which subsequently were spread onto the Langmuir trough under defined conditions.After transfer of aliquots from the surface monolayers to carbon support films the preparations were examined with the electron microscope. The micrographs show threads of various lengths, partly stretched, partly folded in loops, consisting of DNA molecules embedded in a protein envelope.Measurements and calculations of 5900 particles of the complex of Gadus virens-DNA-Albumin, with relatively short threads show a distribution of discontinuous character. If length is plotted against number then it occurs that there are maxima of different lengths of threads. The abscissae of these maxima obey the ratio 1 : 2 : 4 : 8. This holds for longer threads too the maxima of which, however, have smaller ordinate values.

scholarly journals A reassessment of the assay for the asialoglycoprotein receptor and its use in the quantification of receptor distribution in hepatocytes

1986 ◽  
Vol 234 (1) ◽  
pp. 131-137 ◽  
Author(s):  
D A W Grant ◽  
N Kaderbhai
Keyword(s):  
Plasma Membrane ◽  
Bovine Serum Albumin ◽  
Ligand Binding ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Asialoglycoprotein Receptor ◽  
Scatchard Analysis ◽  
Liver Protein ◽  
Binding Studies ◽  
Triton X

The assay for the fucose-binding protein described by Lehrman & Hill [(1983) Methods Enzymol. 98, 309-319] was adapted for the measurement of the asialoglycoprotein receptor in rat liver. The amount of ligand bound to the plasma-membrane-associated or affinity-purified receptor was acutely sensitive to the concentrations of Triton X-100 and NaCl in the assay: 0.02% (v/v) Triton X-100 increased ligand binding to the two preparations by 100% and 40% respectively. Higher concentrations of detergent progressively decreased binding, and in 0.32% Triton X-100 it was about 30% of the value obtained in detergent-free buffer. The addition of increasing concentrations of NaCl to the assay progressively inhibited ligand binding to the membrane-associated receptor, whereas there was a 60% increase in binding to the pure receptor in the presence of 0.1-0.2 M-NaCl. These effects could not be identified in the original assay procedure described by Hudgin, Pricer, Ashwell, Stockert & Morell [(1974) J. Biol. Chem. 249, 5536-5543]. Using optimal assay conditions the binding of 125I-beta-D-galactosyl-bovine serum albumin to both the membrane-associated and purified receptor was inhibited by 50% by 1 nM-beta-D-galactosyl-bovine serum albumin and -asialoorosomucoid and by approx. 100 microM-beta-L-fucosyl-bovine serum albumin, whereas beta-D-galactose, lactose and beta-L-fucose had no effect on ligand binding up to concentrations of 1 mM, 500 microM and 5 mM respectively. KD values of 0.94 and 1.25 nM and Bmax. values of 40 and 1660 pmol of D-galactosyl-bovine serum albumin bound/mg of receptor were obtained for the membrane-bound and purified receptor respectively. Hill-plot analysis of the same data gave slopes of 0.96 and 1.01. Scatchard analysis of saturation-binding studies with other subcellular fractions indicated that the receptor was distributed in the proportions 72:23:2.5:2.5 between total microsomal fractions, plasma membrane, Golgi and canalicular membrane respectively. The receptor was about 1% of the total protein in each compartment and was estimated to be about 0.3% of the total liver protein.

Fragments of bovine serum albumin produced by limited proteolysis. Conformation and ligand binding

Biochemistry ◽  
1975 ◽  
Vol 14 (21) ◽  
pp. 4578-4583 ◽  
Author(s):  
Roberta G. Reed ◽  
Richard C. Feldhoff ◽  
O. L. Clute ◽  
Theodore Peters
Keyword(s):  
Bovine Serum Albumin ◽  
Ligand Binding ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Limited Proteolysis
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