Functional unit size of the charybdotoxin receptor in smooth muscle.
Target inactivation analysis was used to determine the functional size of the charybdotoxin (ChTX) receptor in aortic and tracheal sarcolemmal membrane vesicles. This receptor has previously been shown to be an integral component of the high-conductance Ca2+-activated K+ (Maxi-K) channel in these smooth muscles. Exposure of either bovine aortic or bovine tracheal sarcolemma to high-energy irradiation results in disappearance of 125I-labeled ChTX binding activity as a monoexponential function of radiation dose; from these functions molecular masses of 88 +/- 10 kDa and 89 +/- 6 kDa, respectively, can be calculated. Similar results were obtained from radiation inactivation studies with the detergent-solubilized ChTX receptor from aortic sarcolemmal membranes. The effect of radiation on 125I-labeled ChTX binding is to decrease the number of functional ChTX receptors without affecting the affinity of receptors for the toxin, indicating that radiation is destroying, rather than altering, the binding site. The validity of the radiation inactivation technique in these membrane preparations is supported by data obtained in parallel experiments in which target sizes of the alpha 1 subunit of the L-type Ca2+ channel and 5'-nucleotidase were measured. The molecular masses determined for these entities are in excellent agreement with those expected from previous studies. The present data are discussed in terms of the recently determined subunit composition of the smooth muscle Maxi-K channel. In light of the target size, a single alpha beta subunit heterodimer complex could serve as the ChTX receptor.
Garcia-Calvo
M
M
Department of Membrane Biochemistry and Biophysics, Merck Research Laboratories, Rahway, NJ 07065.
Knaus
H G
HG
Garcia
M L
ML
Kaczorowski
G J
GJ
Kempner
E S
ES
D000818:Animals; D001011:Aorta; D002417:Cattle; D018999:Charybdotoxin; D009130:Muscle, Smooth / Q000378:metabolism* / Q000648:ultrastructure; D015221:Potassium Channels / Q000737:chemistry* / Q000378:metabolism; D012508:Sarcolemma / Q000378:metabolism / Q000648:ultrastructure; D012604:Scorpion Venoms / Q000378:metabolism*; D014132:Trachea
D015221:Potassium Channels; D012604:Scorpion Venoms; C087375:charybdotoxin receptor; D018999:Charybdotoxin