Premium
A universally conserved residue in the SUR1 subunit of the K ATP channel is essential for translating nucleotide binding at SUR1 into channel opening
Author(s) -
de Wet Heidi,
Shimomura Kenju,
Aittoniemi Jussi,
Ahmad Nawaz,
Lafond Mathilde,
Sansom Mark S.P.,
Ashcroft Frances M.
Publication year - 2012
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.2012.236075
Subject(s) - sulfonylurea receptor , atp binding cassette transporter , cyclic nucleotide binding domain , protein subunit , biochemistry , binding site , nucleotide , potassium channel , chemistry , allosteric regulation , biophysics , biology , transporter , enzyme , gene
Key points• The sulphonylurea receptor (SUR1) subunit of the ATP‐sensitive potassium (K ATP ) channel is a member of the ATP‐binding cassette (ABC) protein family. Binding of MgADP to nucleotide‐binding domain 2 (NBD2) is critical for channel activation. • We identified a residue in the SUR1 subunit of the K ATP channel that is essential for translating nucleotide binding to SUR1 into activation of the channel pore. • The ability of ATP to block K ATP channel activity by binding to the Kir6.2 subunit of the channel was also altered by the mutation. This effect was dependent on the integrity of the NBDs of SUR1. This suggests SUR1 also modulates nucleotide inhibition of the channel at Kir6.2. • G1401 in SUR1 is one of 23 residues which are conserved throughout all ABC transporter proteins suggesting that it may have a universal role in coupling substrate binding to protein function.Abstract The sulphonylurea receptor (SUR1) subunit of the ATP‐sensitive potassium (K ATP ) channel is a member of the ATP‐binding cassette (ABC) protein family. Binding of MgADP to nucleotide‐binding domain 2 (NBD2) is critical for channel activation. We identified a residue in NBD2 (G1401) that is fully conserved among ABC proteins and whose functional importance is unknown. Homology modelling places G1401 on the outer surface of the protein, distant from the nucleotide‐binding site. The ATPase activity of purified SUR1‐NBD2‐G1410R (bound to maltose‐binding protein) was slightly inhibited when compared to the wild‐type protein, but its inhibition by MgADP was unchanged, indicating that MgADP binding is not altered. However, MgADP activation of channel activity was abolished. This implies that the G1401R mutation impairs the mechanism by which MgADP binding to NBD2 is translated into opening of the K ATP channel pore. The location of G1401 would be consistent with interaction of this residue with the pore‐forming Kir6.2 subunit. Channel activity in the presence of MgATP reflects the balance between the stimulatory (at SUR1) and inhibitory (at Kir6.2) effects of nucleotides. Mutant channels were 2.5‐fold less sensitive to MgATP inhibition and not activated by MgATP. This suggests that ATP block of the channel is reduced by the SUR1 mutation. Interestingly, this effect was dependent on the functional integrity of the NBDs. These results therefore suggest that SUR1 modulates both nucleotide inhibition and activation of the K ATP channel.