Characterization, Purification, and Affinity Labeling of the Brain [3H]glibenclamide-binding Protein, a Putative Neuronal ATP-regulated K+ Channel

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1988 Dec 1

PMID 3144003

Citations 31

Authors

H Bernardi

M Fosset

M Lazdunski

Affiliations

Soon will be listed here.

Abstract

Sulfonylurea and particularly glibenclamide are potent blockers of ATP-regulated K+ channels in insulin-secreting cells. A very good correlation exists between binding of sulfonylurea to brain and insulinoma cell membranes. The [3H]glibenclamide-binding component from pig brain microsomes was solubilized with digitonin with a complete retention of its properties of interaction with glibenclamide and other sulfonylureas. A four-step purification was achieved that used (i) hydroxylapatite chromatography, (ii and iii) affinity chromatographies on ADP-agarose and wheat germ agglutinin-agarose columns, and (iv) a final chromatographic step on a mixture of AMP-agarose/GMP-agarose/hydroxylapatite. This procedure led to a 2500-fold purification. NaDodSO4/polyacrylamide gel electrophoresis of the purified material in reducing and nonreducing conditions showed that the sulfonylurea-binding component is made of a single major polypeptide chain of Mr 150,000 +/- 10,000. Direct photoaffinity labeling of the receptor with [3H]glibenclamide at different steps of the purification also showed that radioactivity was specifically incorporated into a polypeptide of Mr 150,000 +/- 5000, thus confirming the subunit structure indicated by the purification.

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References

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

REHM H, Lazdunski M . Purification and subunit structure of a putative K+-channel protein identified by its binding properties for dendrotoxin I. Proc Natl Acad Sci U S A. 1988; 85(13):4919-23. PMC: 280549. DOI: 10.1073/pnas.85.13.4919. View

Krueger B, Ratzlaff R, Strichartz G, Blaustein M . Saxitoxin binding to synaptosomes, membranes, and solubilized binding sites from rat brain. J Membr Biol. 1979; 50(3-4):287-310. DOI: 10.1007/BF01868894. View

Merril C, Goldman D, Sedman S, EBERT M . Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981; 211(4489):1437-8. DOI: 10.1126/science.6162199. View

Kaubisch N, Hammer R, Wollheim C, Renold A, Offord R . Specific receptors for sulfonylureas in brain and in a beta-cell tumor of the rat. Biochem Pharmacol. 1982; 31(6):1171-4. DOI: 10.1016/0006-2952(82)90363-x. View

Noma A . ATP-regulated K+ channels in cardiac muscle. Nature. 1983; 305(5930):147-8. DOI: 10.1038/305147a0. View

Lombet A, Lazdunski M . Characterization, solubilization, affinity labeling and purification of the cardiac Na+ channel using Tityus toxin gamma. Eur J Biochem. 1984; 141(3):651-60. DOI: 10.1111/j.1432-1033.1984.tb08241.x. View

Borsotto M, Norman R, Fosset M, Lazdunski M . Solubilization of the nitrendipine receptor from skeletal muscle transverse tubule membranes. Interactions with specific inhibitors of the voltage-dependent Ca2+ channel. Eur J Biochem. 1984; 142(3):449-55. DOI: 10.1111/j.1432-1033.1984.tb08307.x. View

Hansen A . Effect of anoxia on ion distribution in the brain. Physiol Rev. 1985; 65(1):101-48. DOI: 10.1152/physrev.1985.65.1.101. View

10.

Geisen K, Hitzel V, Okomonopoulos R, Punter J, Weyer R, SUMM H . Inhibition of 3H-glibenclamide binding to sulfonylurea receptors by oral antidiabetics. Arzneimittelforschung. 1985; 35(4):707-12. View

11.

Sturgess N, Ashford M, Cook D, Hales C . The sulphonylurea receptor may be an ATP-sensitive potassium channel. Lancet. 1985; 2(8453):474-5. DOI: 10.1016/s0140-6736(85)90403-9. View

12.

Galizzi J, Borsotto M, Barhanin J, Fosset M, Lazdunski M . Characterization and photoaffinity labeling of receptor sites for the Ca2+ channel inhibitors d-cis-diltiazem, (+/-)-bepridil, desmethoxyverapamil, and (+)-PN 200-110 in skeletal muscle transverse tubule membranes. J Biol Chem. 1986; 261(3):1393-7. View

13.

Black A, Dolly J . Two acceptor sub-types for dendrotoxin in chick synaptic membranes distinguishable by beta-bungarotoxin. Eur J Biochem. 1986; 156(3):609-17. DOI: 10.1111/j.1432-1033.1986.tb09621.x. View

14.

Misler S, Falke L, Gillis K, McDaniel M . A metabolite-regulated potassium channel in rat pancreatic B cells. Proc Natl Acad Sci U S A. 1986; 83(18):7119-23. PMC: 386665. DOI: 10.1073/pnas.83.18.7119. View

15.

Dunne M, Petersen O . Intracellular ADP activates K+ channels that are inhibited by ATP in an insulin-secreting cell line. FEBS Lett. 1986; 208(1):59-62. DOI: 10.1016/0014-5793(86)81532-0. View

16.

Kakei M, Kelly R, Ashcroft S, Ashcroft F . The ATP-sensitivity of K+ channels in rat pancreatic B-cells is modulated by ADP. FEBS Lett. 1986; 208(1):63-6. DOI: 10.1016/0014-5793(86)81533-2. View

17.

Levinson S, Duch D, Urban B, Recio-Pinto E . The sodium channel from Electrophorus electricus. Ann N Y Acad Sci. 1986; 479:162-78. DOI: 10.1111/j.1749-6632.1986.tb15568.x. View

18.

Stevens C . Molecular neurobiology. Channel families in the brain. Nature. 1987; 328(6127):198-9. DOI: 10.1038/328198b0. View

19.

Petersen O, Findlay I . Electrophysiology of the pancreas. Physiol Rev. 1987; 67(3):1054-116. DOI: 10.1152/physrev.1987.67.3.1054. View

20.

Schmid-Antomarchi H, De Weille J, Fosset M, Lazdunski M . The antidiabetic sulfonylurea glibenclamide is a potent blocker of the ATP-modulated K+ channel in insulin secreting cells. Biochem Biophys Res Commun. 1987; 146(1):21-5. DOI: 10.1016/0006-291x(87)90684-x. View