scholarly journals Reversible protein affinity-labelling using bromomaleimide-based reagents

2013 ◽  
Vol 11 (15) ◽  
pp. 2408 ◽  
Author(s):  
Ramiz I. Nathani ◽  
Vijay Chudasama ◽  
Chris P. Ryan ◽  
Paul R. Moody ◽  
Rachel E. Morgan ◽  
...  
Keyword(s):  
Protein Affinity ◽  
Affinity Labelling

First protein affinity application of Cu2+-bound pure inorganic nanoflowers

2021 ◽  
Author(s):  
Burcu Önal ◽  
Ömür Acet ◽  
Volha Dzmitruk ◽  
Inessa Halets-Bui ◽  
Dzmitry Shcharbin ◽  
...  
Keyword(s):  
Protein Affinity

scholarly journals Escherichia coli dnaB protein. Affinity chromatography on immobilized nucleotides

1978 ◽  
Vol 253 (16) ◽  
pp. 5847-5851 ◽  
Author(s):  
E. Lanka ◽  
C. Edelbluth ◽  
M. Schlicht ◽  
H. Schuster
Keyword(s):  
Escherichia Coli ◽  
Affinity Chromatography ◽  
Protein Affinity

scholarly journals Structures of the somatotropin receptor and prolactin receptor on rat hepatocytes characterized by affinity labelling

1984 ◽  
Vol 220 (2) ◽  
pp. 361-369 ◽  
Author(s):  
K Yamada ◽  
D B Donner
Keyword(s):  
Binding Sites ◽  
Prolactin Receptor ◽  
Rat Hepatocytes ◽  
Membrane Receptors ◽  
Bovine Somatotropin ◽  
Male Rats ◽  
Disulphide Bonds ◽  
Low Concentrations ◽  
Affinity Labelling ◽  
Structural Descriptions

Human somatotropin competed for 125I-human somatotropin binding to hepatocytes from female or male rats. Bovine somatotropin and prolactin each inhibited part, but not all, of the uptake of 125I-human somatotropin. The binding of 125I-prolactin was inhibited by human somatotropin and prolactin, but not by bovine somatotropin. Bovine somatotropin and human somatotropin, but not prolactin, competed for 125I-bovine somatotropin binding sites. 125I-labelled hormones were covalently coupled to membrane receptors with higher efficiency on hepatocytes from female than from male rats, allowing structural descriptions of lactogenic and somatogenic binding sites that had not been possible previously. Disuccinimidyl suberate covalently coupled 125I-human somatotropin into saturable complexes of Mr 300 000, 220 000, 130 000, 65 000 and 50 000. Bovine somatotropin inhibited the incorporation of 125I-human somatotropin into complexes of Mr 300 000, 220 000 and 130 000, whereas low concentrations of prolactin competed for incorporation into the 65 000- and 50 000-Mr species. 125I-bovine somatotropin was incorporated into complexes of Mr 300 000, 220 000 and 130 000. Human somatotropin and bovine somatotropin, but not prolactin, inhibited the production of these complexes. 125I-prolactin binding produced complexes of Mr 65 000 and 50 000. Native prolactin and human somatotropin, but not bovine somatotropin, inhibited uptake of 125I-prolactin into these species. Thus direct affinity labelling, as well as competition for covalent coupling, suggests that the 300 000-, 220 000- and 130 000-Mr species are components of the somatotropin receptor and that the 65 000- and 50 000-Mr complexes result from hormone binding to the prolactin receptor. By subtracting the Mr of prolactin, it was calculated that the hormone was bound to species of Mr 43 000 and 28 000. These Mr values were not affected by reduction of solubilized membranes, suggesting that the structure of the prolactin receptor is not stabilized by interchain disulphide bonds between subunits. Subtracting the Mr of somatotropin from somatogenic complexes indicated that the hormone had bound to species of Mr 280 000, 200 000 and 100 000. The 300 000- and 220 000-Mr complexes were not isolated from reduced membranes, whereas the amount of the 130 000-Mr species was augmented. These observations could suggest that a major component of the somatotropin receptor is a trimeric aggregate in which some subunits are retained in a larger complex by interchain disulphide bonds.

Development of photo-affinity labelling reagents for the thrombin receptor

1995 ◽  
Vol 6 (2) ◽  
pp. 179
Author(s):  
Carla Chinni ◽  
Tony Johnson ◽  
Edward J. Duffy ◽  
Stuart R. Stone
Keyword(s):  
Thrombin Receptor ◽  
Affinity Labelling

Affinity labelling of estradiol receptor by ferrocenyl tagging of estradiol 17α-position

1990 ◽  
pp. 837-839 ◽  
Author(s):  
Anne Vessiēres ◽  
Colette Vaillant ◽  
Michel Gruselle ◽  
Dominique Vichard ◽  
Gérard Jaouen
Keyword(s):  
Estradiol Receptor ◽  
Affinity Labelling

scholarly journals Affinity labelling of the binding site of rabbit antibody. Evidence for the involvement of the hypervariable regions of the heavy chain

1974 ◽  
Vol 139 (1) ◽  
pp. 135-149 ◽  
Author(s):  
Christopher E. Fisher ◽  
Elizabeth M. Press
Keyword(s):  
Binding Site ◽  
Heavy Chain ◽  
Binding Sites ◽  
Light Chains ◽  
Rabbit Antibody ◽  
Heavy Chains ◽  
Group 4 ◽  
Hypervariable Regions ◽  
Affinity Labelling ◽  
Antibody Complex

The binding sites of rabbit antibodies with affinity for the haptenic group 4-azido-2-nitrophenyl-lysine have been specifically labelled by photolysis of the hapten–antibody complex. The extent of covalent labelling was 0.5–0.9mol of hapten bound/mol of antibody and, by using an immunoadsorbent, antibody with 1.3mol of hapten/mol was obtained. The antibody was specifically labelled in the binding site and the ratio of labelling of heavy and light chains was in the range 3.3–5.0. The labelled heavy chains were cleaved by CNBr treatment and after reduction and alkylation of the intrachain bonds, were digested with trypsin. Evidence is presented that two regions of the heavy chain, positions 29–34 and 95–114, together contain about 80% of the label on the heavy chain; these two regions respectively include two of the hypervariable regions of rabbit heavy chain.

Sign in / Sign up

Export Citation Format

Share Document