Cryogenic solvents inhibit the binding of the Escherichia coli heat-stable enterotoxin to intestinal brush border membranes.
The cryogenic solvents ethylene glycol, glycerol, dimethyl sulfoxide (Me2SO) and dimethylformamide (Me2FM), with increasing potency, produced concentration-dependent inhibition of the binding of the Escherichia coli heat-stable enterotoxin (STa) to pig intestinal brush border membranes. Inhibition increased with time, and both Me2SO and Me2FM appeared to decrease both the affinity of the STa receptor and the effective receptor number. Both solvents stimulated the release of previously bound 125I-STa (Me2FM > Me2SO), 3 M Me2FM inducing 93% release by 120 min. These effects were reversible, and preincubation of membranes with up to 3 M Me2SO or Me2FM at 37 degrees C for 30 min, followed by washing, did not alter subsequent 125I-STa binding. Also, 125I-STa released from membranes by 3 M Me2FM was shown to rebind to the membranes after 10-fold dilution of Me2FM. Since pretreating membranes with the thiol reagent p-chloromercuribenzenesulfonate had no effect on the release of bound 125I-STa by Me2SO or Me2FM, and since neither of these can reduce disulfide bonds, the formation of mixed disulfides between STa and receptor is unlikely. Me2SO inhibition of 125I-STa binding was greater with membranes than with a partially purified receptor preparation, which may result from the substitution of detergent for the phospholipid normally associated with the receptor(s).
Katwa
L C
LC
John M. Dalton Research Center, Department of Molecular Microbiology, University of Missouri-Columbia 65211.
Parker
C D
CD
Dybing
J K
JK
White
A A
AA
D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology; D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism*; D001667:Binding, Competitive; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004126:Dimethylformamide / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004768:Enterotoxins / Q000378:metabolism*; D029968:Escherichia coli Proteins; D019855:Ethylene Glycol; D005026:Ethylene Glycols / Q000494:pharmacology; D005990:Glycerol / Q000494:pharmacology; D007583:Jejunum / Q000378:metabolism*; D008871:Microvilli / Q000378:metabolism*; D013552:Swine; D013997:Time Factors
D001427:Bacterial Toxins; D003451:Cryoprotective Agents; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D005026:Ethylene Glycols; C034151:heat stable toxin (E coli); D002731:4-Chloromercuribenzenesulfonate; D004126:Dimethylformamide; D019855:Ethylene Glycol; D005990:Glycerol; D004121:Dimethyl Sulfoxide