Guanine nucleotide regulation of B2 kinin receptors. Time-dependent formation of a guanine nucleotide-sensitive receptor state from which [3H]bradykinin dissociates slowly
(1990) In Journal of Biological Chemistry 265(17). p.9621-9627- Abstract
We have examined the binding of [3H]bradykinin to bovine myometrial membranes and assessed its sensitivity to guanine nucleotides. Total binding displayed a typical B2 kinin receptor specificity. However, saturation binding isotherms were resolved into at least two components with K(D) values of 8 pM (45%) and 378 pM (55%). Low affinity binding exhibited relatively rapid rates of association (k(obs) = 1.40 x 10-2 s-1) and dissociation (k-1 = 3.82 x 10-3 s-1), while high affinity binding exhibited considerably slower rates (k(obs) = 9.52 x 10-4 s-1 and k-1 = 4.43 x 10-5 s-1). Pre-equilibrium dissociation kinetics... (More)
We have examined the binding of [3H]bradykinin to bovine myometrial membranes and assessed its sensitivity to guanine nucleotides. Total binding displayed a typical B2 kinin receptor specificity. However, saturation binding isotherms were resolved into at least two components with K(D) values of 8 pM (45%) and 378 pM (55%). Low affinity binding exhibited relatively rapid rates of association (k(obs) = 1.40 x 10-2 s-1) and dissociation (k-1 = 3.82 x 10-3 s-1), while high affinity binding exhibited considerably slower rates (k(obs) = 9.52 x 10-4 s-1 and k-1 = 4.43 x 10-5 s-1). Pre-equilibrium dissociation kinetics revealed that formation of high affinity binding was characterized as a time-dependent accumulation of the slow dissociation rate at the expense of at least one other more rapid dissociation rate. In the presence of 10 μM guanyl-5'-yl imidodiphosphate (Gpp(NH)p), at least two binding components were resolved with K(D) values of 37 pM (12%) and 444 pM (88%). Gpp(NH)p apparently specifically perturbed high affinity binding by completely preventing the accumulation of the slow dissociation phase. Instead, two more rapid dissociation rates (k-1 = 8.53 x 10-3 s-1 and 4.43 x 10-4 s-1) were observed. These results suggest that [3H]bradykinin interacts with at least two B2 kinin receptor-like binding sites in bovine myometrial membranes. A three-state model for the guanine nucleotide-sensitive agonist interaction with the high affinity binding sites is proposed.
(Less)
- author
- Leeb-Lundberg, L. M.F. LU and Mathis, S. A.
- publishing date
- 1990-07-09
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 265
- issue
- 17
- pages
- 7 pages
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- pmid:2161838
- scopus:0025324527
- ISSN
- 0021-9258
- language
- English
- LU publication?
- no
- id
- abc80680-60b1-4cbb-a532-ee13b06997bb
- alternative location
- http://www.jbc.org/content/265/17/9621.abstract
- date added to LUP
- 2019-06-04 14:17:57
- date last changed
- 2025-04-04 15:16:57
@article{abc80680-60b1-4cbb-a532-ee13b06997bb,
abstract = {{<p>We have examined the binding of [<sup>3</sup>H]bradykinin to bovine myometrial membranes and assessed its sensitivity to guanine nucleotides. Total binding displayed a typical B2 kinin receptor specificity. However, saturation binding isotherms were resolved into at least two components with K(D) values of 8 pM (45%) and 378 pM (55%). Low affinity binding exhibited relatively rapid rates of association (k(obs) = 1.40 x 10<sup>-2</sup> s<sup>-1</sup>) and dissociation (k<sub>-1</sub> = 3.82 x 10<sup>-3</sup> s<sup>-1</sup>), while high affinity binding exhibited considerably slower rates (k(obs) = 9.52 x 10<sup>-4</sup> s<sup>-1</sup> and k<sub>-1</sub> = 4.43 x 10<sup>-5</sup> s<sup>-1</sup>). Pre-equilibrium dissociation kinetics revealed that formation of high affinity binding was characterized as a time-dependent accumulation of the slow dissociation rate at the expense of at least one other more rapid dissociation rate. In the presence of 10 μM guanyl-5'-yl imidodiphosphate (Gpp(NH)p), at least two binding components were resolved with K(D) values of 37 pM (12%) and 444 pM (88%). Gpp(NH)p apparently specifically perturbed high affinity binding by completely preventing the accumulation of the slow dissociation phase. Instead, two more rapid dissociation rates (k<sub>-1</sub> = 8.53 x 10<sup>-3</sup> s<sup>-1</sup> and 4.43 x 10<sup>-4</sup> s<sup>-1</sup>) were observed. These results suggest that [<sup>3</sup>H]bradykinin interacts with at least two B2 kinin receptor-like binding sites in bovine myometrial membranes. A three-state model for the guanine nucleotide-sensitive agonist interaction with the high affinity binding sites is proposed.</p>}},
author = {{Leeb-Lundberg, L. M.F. and Mathis, S. A.}},
issn = {{0021-9258}},
language = {{eng}},
month = {{07}},
number = {{17}},
pages = {{9621--9627}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
series = {{Journal of Biological Chemistry}},
title = {{Guanine nucleotide regulation of B2 kinin receptors. Time-dependent formation of a guanine nucleotide-sensitive receptor state from which [<sup>3</sup>H]bradykinin dissociates slowly}},
url = {{http://www.jbc.org/content/265/17/9621.abstract}},
volume = {{265}},
year = {{1990}},
}