Four novel splice variants of sulfonylurea receptor 1.
ATP-sensitive K(+) (K(ATP)) channels are composed of pore-forming Kir6.x subunits and regulatory sulfonylurea receptor (SUR) subunits. SURs are ATP-binding cassette proteins with two nucleotide-binding folds (NBFs) and binding sites for sulfonylureas, like glibenclamide, and for channel openers. Here we report the identification and functional characterization of four novel splice forms of guinea pig SUR1. Three splice forms originate from alternative splicing of the region coding for NBF1 and lack exons 17 (SUR1Delta17), 19 (SUR1Delta19), or both (SUR1Delta17Delta19). The fourth (SUR1C) is a COOH-terminal SUR1-fragment formed by exons 31-39 containing the last two transmembrane segments and the COOH terminus of SUR1. RT-PCR analysis showed that these splice forms are expressed in several tissues with strong expression of SUR1C in cardiomyocytes. Confocal microscopy using enhanced green fluorescent protein-tagged SUR or Kir6.x did not provide any evidence for involvement of these splice forms in the mitochondrial K(ATP) channel. Only SUR1 and SUR1Delta17 showed high-affinity binding of glibenclamide (K(d) approximately 2 nM in the presence of 1 mM ATP) and formed functional K(ATP) channels upon coexpression with Kir6.2.
Hambrock
Annette
A
Institute of Pharmacology, Tübingen University, 72074 Tübingen, Germany.
Preisig-Müller
Regina
R
Russ
Ulrich
U
Piehl
Anke
A
Hanley
Peter J
PJ
Ray
John
J
Daut
Jürgen
J
Quast
Ulrich
U
Derst
Christian
C
D018528:ATP-Binding Cassette Transporters; D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D001483:Base Sequence / Q000235:genetics; D019556:COS Cells; D002458:Cell Fractionation; D002460:Cell Line; D004274:DNA, Recombinant / Q000235:genetics*; D004594:Electrophysiology; D005905:Glyburide / Q000378:metabolism; D006168:Guinea Pigs; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism; D015221:Potassium Channels / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D024661:Potassium Channels, Inwardly Rectifying; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011955:Receptors, Drug / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D064233:Sulfonylurea Receptors; D014018:Tissue Distribution
C577507:ABCC8 protein, human; D018528:ATP-Binding Cassette Transporters; C577509:Abcc8 protein, rat; D004274:DNA, Recombinant; D008565:Membrane Proteins; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D020033:Protein Isoforms; D011955:Receptors, Drug; D064233:Sulfonylurea Receptors; C117780:mitochondrial K(ATP) channel; D005905:Glyburide