Binding and hydrodynamic properties of muscarinic receptor subtypes solubilized in 3-(3-cholamidopropyl)dimethylammonio-2-hydroxy-1-propanesulfonate.
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.
Poyner
D R
DR
Division of Physical Biochemistry, National Institute for Medical Research, London, UK.
Birdsall
N J
NJ
Curtis
C
C
Eveleigh
P
P
Hulme
E C
EC
Pedder
E K
EK
Wheatley
M
M
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002540:Cerebral Cortex / Q000378:metabolism; D002793:Cholic Acids; D003902:Detergents; D019204:GTP-Binding Proteins / Q000378:metabolism; D006165:Guanylyl Imidodiphosphate / Q000378:metabolism; D007765:Lacrimal Apparatus / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D019832:N-Methylscopolamine; D010095:Oxotremorine / Q000378:metabolism; D051381:Rats; D011976:Receptors, Muscarinic / Q000145:classification / Q000378:metabolism*; D012602:Scopolamine Derivatives / Q000378:metabolism; D012995:Solubility
D002793:Cholic Acids; D003902:Detergents; D011976:Receptors, Muscarinic; D012602:Scopolamine Derivatives; D006165:Guanylyl Imidodiphosphate; D010095:Oxotremorine; C048531:chapso; D019204:GTP-Binding Proteins; D019832:N-Methylscopolamine