Binding of bumetanide to microsomes from optic ganglia of the squid, Loligo pealei

A. A. Altamirano, Bruns Watts, J. M. Russell

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Saturable high-affinity binding of [3H]bumetanide [dissociation constant (K(D)) = 80 nM] was measured in microsomal membranes prepared from squid optic ganglia. Under control conditions, the maximal specific binding of labeled bumetanide (B(max)) was ~6-7 pmol/mg protein. Binding had a higher relative affinity for bumetanide than for furosemide and dependend on the presence of Cl- and K+, but not Na+, in the incubation media. In the case of K+, [3H]-bumetanide binding was half-saturated at [K+] = 100 mM. The Cl- effect was biphasic. At [Cl-] between 0 and 150 mM, [3H]-bumetanide binding increased with increasing [Cl-]. However, when [Cl-] was increased above 150 mM, [3H]bumetanide binding was progressively reduced. ATP acted as a nonessential activator [mean affinity constant (K0.5) ~1 μM] of the ion-dependent [3H]bumetanide binding by increasing the apparent binding capacity. The activation by ATP did not require Mg2+. Other adenosine analogues also stimulated the binding of bumetanide.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume258
Issue number5 27-5
StatePublished - 1990

Fingerprint

Loligo
Bumetanide
Decapodiformes
Microsomes
Ganglia
Optics
Adenosine Triphosphate
Furosemide
Adenosine
Chemical activation
Ions
Membranes

Keywords

  • adenosine 5'-trisphosphate
  • loop diuretics
  • sodium-potassium-chloride cotransport

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Binding of bumetanide to microsomes from optic ganglia of the squid, Loligo pealei. / Altamirano, A. A.; Watts, Bruns; Russell, J. M.

In: American Journal of Physiology - Cell Physiology, Vol. 258, No. 5 27-5, 1990.

Research output: Contribution to journalArticle

@article{dba6ec64b2a44351ac3ad92d0a68306e,
title = "Binding of bumetanide to microsomes from optic ganglia of the squid, Loligo pealei",
abstract = "Saturable high-affinity binding of [3H]bumetanide [dissociation constant (K(D)) = 80 nM] was measured in microsomal membranes prepared from squid optic ganglia. Under control conditions, the maximal specific binding of labeled bumetanide (B(max)) was ~6-7 pmol/mg protein. Binding had a higher relative affinity for bumetanide than for furosemide and dependend on the presence of Cl- and K+, but not Na+, in the incubation media. In the case of K+, [3H]-bumetanide binding was half-saturated at [K+] = 100 mM. The Cl- effect was biphasic. At [Cl-] between 0 and 150 mM, [3H]-bumetanide binding increased with increasing [Cl-]. However, when [Cl-] was increased above 150 mM, [3H]bumetanide binding was progressively reduced. ATP acted as a nonessential activator [mean affinity constant (K0.5) ~1 μM] of the ion-dependent [3H]bumetanide binding by increasing the apparent binding capacity. The activation by ATP did not require Mg2+. Other adenosine analogues also stimulated the binding of bumetanide.",
keywords = "adenosine 5'-trisphosphate, loop diuretics, sodium-potassium-chloride cotransport",
author = "Altamirano, {A. A.} and Bruns Watts and Russell, {J. M.}",
year = "1990",
language = "English (US)",
volume = "258",
journal = "American Journal of Physiology - Endocrinology and Metabolism",
issn = "0193-1849",
publisher = "American Physiological Society",
number = "5 27-5",

}

TY - JOUR

T1 - Binding of bumetanide to microsomes from optic ganglia of the squid, Loligo pealei

AU - Altamirano, A. A.

AU - Watts, Bruns

AU - Russell, J. M.

PY - 1990

Y1 - 1990

N2 - Saturable high-affinity binding of [3H]bumetanide [dissociation constant (K(D)) = 80 nM] was measured in microsomal membranes prepared from squid optic ganglia. Under control conditions, the maximal specific binding of labeled bumetanide (B(max)) was ~6-7 pmol/mg protein. Binding had a higher relative affinity for bumetanide than for furosemide and dependend on the presence of Cl- and K+, but not Na+, in the incubation media. In the case of K+, [3H]-bumetanide binding was half-saturated at [K+] = 100 mM. The Cl- effect was biphasic. At [Cl-] between 0 and 150 mM, [3H]-bumetanide binding increased with increasing [Cl-]. However, when [Cl-] was increased above 150 mM, [3H]bumetanide binding was progressively reduced. ATP acted as a nonessential activator [mean affinity constant (K0.5) ~1 μM] of the ion-dependent [3H]bumetanide binding by increasing the apparent binding capacity. The activation by ATP did not require Mg2+. Other adenosine analogues also stimulated the binding of bumetanide.

AB - Saturable high-affinity binding of [3H]bumetanide [dissociation constant (K(D)) = 80 nM] was measured in microsomal membranes prepared from squid optic ganglia. Under control conditions, the maximal specific binding of labeled bumetanide (B(max)) was ~6-7 pmol/mg protein. Binding had a higher relative affinity for bumetanide than for furosemide and dependend on the presence of Cl- and K+, but not Na+, in the incubation media. In the case of K+, [3H]-bumetanide binding was half-saturated at [K+] = 100 mM. The Cl- effect was biphasic. At [Cl-] between 0 and 150 mM, [3H]-bumetanide binding increased with increasing [Cl-]. However, when [Cl-] was increased above 150 mM, [3H]bumetanide binding was progressively reduced. ATP acted as a nonessential activator [mean affinity constant (K0.5) ~1 μM] of the ion-dependent [3H]bumetanide binding by increasing the apparent binding capacity. The activation by ATP did not require Mg2+. Other adenosine analogues also stimulated the binding of bumetanide.

KW - adenosine 5'-trisphosphate

KW - loop diuretics

KW - sodium-potassium-chloride cotransport

UR - http://www.scopus.com/inward/record.url?scp=0025331157&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025331157&partnerID=8YFLogxK

M3 - Article

C2 - 2333985

AN - SCOPUS:0025331157

VL - 258

JO - American Journal of Physiology - Endocrinology and Metabolism

JF - American Journal of Physiology - Endocrinology and Metabolism

SN - 0193-1849

IS - 5 27-5

ER -