Binding and hydrodynamic properties of muscarinic receptor subtypes solubilized in 3-(3-cholamidopropyl)dimethylammonio-2-hydroxy-1-propanesulfonate

David R. Poyner, N.J. Birdsall, C. Curtis, P. Eveleigh, E.C. Hulme, E.K. Pedder, Mark Wheatley

Research output: Contribution to journalArticle

Abstract

The muscarinic receptor from the cerebral cortex, heart, and lacrimal gland can be solubilized in the zwitterionic detergent 3-(3-cholamidopropyl)dimethylammonio-2-hydroxy-1-propane sulfonate (CHAPSO) with retention of high affinity [3H]N-methyls-copolamine binding. However, in this detergent there are significant differences in the binding properties of the receptors, compared with those observed in membranes and digitonin solution. Some agents retain a degree of selectivity. In the heart and cortex, agonists can bind with high affinity to a receptor-GTP-binding protein complex. A second, lower affinity, agonist binding state is also present, which resembles a class of sites seen in membranes but not in digitonin solution. The high affinity agonist binding state has been resolved from the lower affinity state on sucrose density gradient centrifugation. Hydrodynamic analysis suggests that the high affinity state is approximately 110,000 Da larger than the lower affinity state. The binding properties of the receptor in CHAPSO can be altered to those seen in digitonin by exchanging detergents after CHAPSO solubilization.
Original languageEnglish
Pages (from-to)420-429
Number of pages10
JournalMolecular Pharmacology
Volume36
Issue number3
Publication statusPublished - 26 Oct 1989

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Digitonin
Muscarinic Receptors
Hydrodynamics
Detergents
Lacrimal Apparatus
Membranes
Density Gradient Centrifugation
GTP-Binding Proteins
Cerebral Cortex
Sucrose
propylsulfonic acid
chapso

Keywords

  • animals
  • cell membranecentrifugation
  • density gradient
  • cerebral cortex
  • cholic acids
  • detergents
  • GTP-binding proteins
  • guanylyl imidodiphosphate
  • lacrimal apparatus myocardium
  • N-Methylscopolamine
  • oxotremorine
  • rats
  • receptors
  • muscarinic scopolamine
  • derivatives solubility

Cite this

Poyner, David R. ; Birdsall, N.J. ; Curtis, C. ; Eveleigh, P. ; Hulme, E.C. ; Pedder, E.K. ; Wheatley, Mark. / Binding and hydrodynamic properties of muscarinic receptor subtypes solubilized in 3-(3-cholamidopropyl)dimethylammonio-2-hydroxy-1-propanesulfonate. In: Molecular Pharmacology. 1989 ; Vol. 36, No. 3. pp. 420-429.
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abstract = "The muscarinic receptor from the cerebral cortex, heart, and lacrimal gland can be solubilized in the zwitterionic detergent 3-(3-cholamidopropyl)dimethylammonio-2-hydroxy-1-propane sulfonate (CHAPSO) with retention of high affinity [3H]N-methyls-copolamine binding. However, in this detergent there are significant differences in the binding properties of the receptors, compared with those observed in membranes and digitonin solution. Some agents retain a degree of selectivity. In the heart and cortex, agonists can bind with high affinity to a receptor-GTP-binding protein complex. A second, lower affinity, agonist binding state is also present, which resembles a class of sites seen in membranes but not in digitonin solution. The high affinity agonist binding state has been resolved from the lower affinity state on sucrose density gradient centrifugation. Hydrodynamic analysis suggests that the high affinity state is approximately 110,000 Da larger than the lower affinity state. The binding properties of the receptor in CHAPSO can be altered to those seen in digitonin by exchanging detergents after CHAPSO solubilization.",
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Binding and hydrodynamic properties of muscarinic receptor subtypes solubilized in 3-(3-cholamidopropyl)dimethylammonio-2-hydroxy-1-propanesulfonate. / Poyner, David R.; Birdsall, N.J.; Curtis, C.; Eveleigh, P.; Hulme, E.C.; Pedder, E.K.; Wheatley, Mark.

In: Molecular Pharmacology, Vol. 36, No. 3, 26.10.1989, p. 420-429.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Binding and hydrodynamic properties of muscarinic receptor subtypes solubilized in 3-(3-cholamidopropyl)dimethylammonio-2-hydroxy-1-propanesulfonate

AU - Poyner, David R.

AU - Birdsall, N.J.

AU - Curtis, C.

AU - Eveleigh, P.

AU - Hulme, E.C.

AU - Pedder, E.K.

AU - Wheatley, Mark

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N2 - The muscarinic receptor from the cerebral cortex, heart, and lacrimal gland can be solubilized in the zwitterionic detergent 3-(3-cholamidopropyl)dimethylammonio-2-hydroxy-1-propane sulfonate (CHAPSO) with retention of high affinity [3H]N-methyls-copolamine binding. However, in this detergent there are significant differences in the binding properties of the receptors, compared with those observed in membranes and digitonin solution. Some agents retain a degree of selectivity. In the heart and cortex, agonists can bind with high affinity to a receptor-GTP-binding protein complex. A second, lower affinity, agonist binding state is also present, which resembles a class of sites seen in membranes but not in digitonin solution. The high affinity agonist binding state has been resolved from the lower affinity state on sucrose density gradient centrifugation. Hydrodynamic analysis suggests that the high affinity state is approximately 110,000 Da larger than the lower affinity state. The binding properties of the receptor in CHAPSO can be altered to those seen in digitonin by exchanging detergents after CHAPSO solubilization.

AB - The muscarinic receptor from the cerebral cortex, heart, and lacrimal gland can be solubilized in the zwitterionic detergent 3-(3-cholamidopropyl)dimethylammonio-2-hydroxy-1-propane sulfonate (CHAPSO) with retention of high affinity [3H]N-methyls-copolamine binding. However, in this detergent there are significant differences in the binding properties of the receptors, compared with those observed in membranes and digitonin solution. Some agents retain a degree of selectivity. In the heart and cortex, agonists can bind with high affinity to a receptor-GTP-binding protein complex. A second, lower affinity, agonist binding state is also present, which resembles a class of sites seen in membranes but not in digitonin solution. The high affinity agonist binding state has been resolved from the lower affinity state on sucrose density gradient centrifugation. Hydrodynamic analysis suggests that the high affinity state is approximately 110,000 Da larger than the lower affinity state. The binding properties of the receptor in CHAPSO can be altered to those seen in digitonin by exchanging detergents after CHAPSO solubilization.

KW - animals

KW - cell membranecentrifugation

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KW - cerebral cortex

KW - cholic acids

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KW - GTP-binding proteins

KW - guanylyl imidodiphosphate

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KW - N-Methylscopolamine

KW - oxotremorine

KW - rats

KW - receptors

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