Design of Angiotensin II Derivatives Suitable for Indirect Affinity Techniques: Potential Applications to Receptor Studies
- 1 January 1988
- journal article
- research article
- Published by Taylor & Francis in Journal of Receptor Research
- Vol. 8 (1-4) , 295-309
- https://doi.org/10.3109/10799898809048994
Abstract
The design of angiotensin II (A II)-derived probes suitable for indirect affinity techniques is presented. Biotin or dinitrophenyl moieties have been added at the N-terminus of A II, through aminohexanoic acid as spacer arm, to generate (6-biotinylamido)-hexanoyl-All (Bio-Ahx-All) and dinitrophenyl- aminohexanoyl-All (Dnp-Ahx-All). Monoiodinated and highly labeled radioiodinated forms of these probes have been prepared. The two bifunctional ligands displayed high affinities for rat liver A II receptors (Kd values in the nanomolar range) and their secondary acceptors: streptavidin and monoclonal anti-Dnp antibodies respectively. Bio-Ahx-All and Dnp-Ahx-All behaved as agonists on several All-sensitive systems. Based on these structural assessments, the parent photoactivable azido probe: Bio-Ahx-(Ala1,Phe(4N3)8)A II. A II was synthesized and proved to possess similar biological properties than the non-azido compound. The hepatic A II receptor could be covalently labeled by the radioiodinated probe, with a particularly high yield (15-20%); SDS-polyacrylamide gel electrophoresis of solubilized complexes revealed specific labeling of a 65 Kdaltons binding unit, in agreement with previous data obtained with other azido All-derived compounds. The potential applications of these probes are: i) receptor purification by combination of its photoaffinity labeling and adsorption of biotin-tagged solubilized hormone-receptor complexes on avidin gels. ii) cell labeling and sorting. iii) histochemical receptor visualization.Keywords
This publication has 23 references indexed in Scilit:
- Affinity purification of the opioid receptor in NG 108‐15 cells using an avidin‐biotin system with a novel elution methodFEBS Letters, 1986
- Cytochemical Studies of Corticotropin-Releasing Factor (CRF) Receptors in Anterior Lobe Corticotropes: Binding, Glucocorticoid Regulation, and Endocytosis of [Biotinyl-Ser1]CRF*Endocrinology, 1986
- Radioactive probes for adrenocorticotropic hormone receptorsBiochemistry, 1986
- Receptor Binding Activities of Biotinylated Derivatives of β‐Nerve Growth FactorJournal of Neurochemistry, 1986
- Avidin-biotin affinity chromatography: application to the isolation of human placental insulin receptor.Proceedings of the National Academy of Sciences, 1984
- Binding affinities to rat brain synaptosomes – synthesis of biotinylated analogues of Substance PInternational Journal of Peptide and Protein Research, 1984
- Solubization of angiotensin II receptors in rat glomeruliFEBS Letters, 1984
- The avidin-biotin complex in immunologyImmunology Today, 1984
- Solubilization and characterization of an angiotensin II binding protein from liverEuropean Journal of Biochemistry, 1983
- Solubilization of Adrenal Angiotensin II Receptors with a Zwitterionic DetergentJournal of Receptor Research, 1983