Adenosine 5'-monophosphate is a selective inhibitor of the brown adipocyte nonselective cation channel

Abstract

Calcium-activated nonselective cation channels (NSC(Ca)) in brown adipocytes are inhibited by several nucleotides acting on the cytosolic side of the membrane. We used excised inside-out patches from rat brown adipocytes to identify important nucleotide structures for NSC-channel inhibition. We found that 100 microM 5'-AMP inhibited NSC-channel activity more than did ATP or ADP. Adenosine was a weak inhibitor, whereas adenine and ribose-5-phosphate had no effect. The channel activity was effectively blocked by 10 microM AMP, but it was unaffected by 10 microM cAMP, CMP, GMP, IMP, TMP or UMP. Dose-response studies yielded IC(50)-values of 4 microM for AMP and 32 microM for cAMP. dAMP was as effective as AMP, but all 5'-phosphate group modifications on AMP dramatically lowered the inhibitory effect. 10 microM of the AMP precursor adenylosuccinate weakly inhibited the channel activity. An increase in AMP concentration from 1 to 10 microM shifted the EC(50) for Ca(2+) activation almost 1 order of magnitude; a Schild plot analysis yielded a K(B) value of 0.3 microM for AMP. We conclude that AMP is the most efficacious endogenous nucleotide inhibitor of the brown adipocyte nonselective cation channel (NSC(Ca/AMP)) yet identified and that there is functional competition between Ca(2+) and AMP. The brown adipocyte NSC(Ca/AMP) thus appears to be functionally different from the NSC(Ca,PKA) in the exocrine pancreas and the NSC-(Ca,cAMP) in the endocrine pancreas, but similar to the NSC(Ca/AMP) in the endocrine pancreas.