Korean J Physiol Pharmacol.  1997 Feb;1(1):19-25.

Enhanced coupling of M-1 muscarinic receptors to activation of phospholipase C upon mutation of a transposed amino acid triplet repeat

• Lee SY
• Sung KW
• Kim ON
• Lee SB

Abstract

The C-terminus ends of the second putative transmembrane domains of both M-1 and M-2 Muscarinic receptors contain a triplet of amino acid residues consisting of leucine (L), tyrosine (Y) and threonine (T). This triplet is repeated as LYT-TYL in M-1 receptors at the interface between the second transmembrane domain and the first extracellular loop. Interestingly, however, it is repeated in a transposed fashion (LYT-LYT) in the sequence Of M-2 receptors. In our previous work, we investigated the possible significance of this unique sequence diversity for determining the distinct differential receptor function at the two receptor subtypes. However, we found mutation of the LYTTYL sequence of M-1 receptors to the corresponding M-2 receptor LYTLYT sequence demonstrated markedly enhanced the stimulation of phosphoinositide (PI) hydrolysis by carbachol without a change in its coupling to increased cyclic AMP formation. In this work, thus, the enhanced stimulation of PI hydrolysis in the LYTLYT M-1 receptor mutant was further investigated. The stimulation of PI hydrolysis by carbachol was enhanced in the mutant M-1 receptor, and this change was not due to alterations in the rate of receptor desensitization or sequestration. The observed larger response to carbachol at mutant M-1 receptors was also not due to an artifact resulting from selection of CHO cells which express higher levels of G-proteins or phospholipase C. Our data suggest that although the LYTTYL sequence in M-1 muscarinic receptors is not involved in determining receptor pharmacology, mutation of the sequence enhanced the coupling of M-1 receptors to the stimulation of phospholipase C.

Keyword

M1 muscarinic receptors; Phospholipase C; Site-mutagenesis

MeSH Terms

Animals
Artifacts
Carbachol
CHO Cells
Cricetinae
Cyclic AMP
GTP-Binding Proteins
Humans
Hydrolysis
Leucine
Pharmacology
Phospholipases*
Receptors, Muscarinic*
Threonine
Trinucleotide Repeats*
Triplets*
Type C Phospholipases*
Tyrosine
Carbachol
Cyclic AMP
GTP-Binding Proteins
Leucine
Phospholipases
Receptors, Muscarinic
Threonine
Type C Phospholipases
Tyrosine