Consensus residues at the acetylcholine binding site of cholinergic proteins.
The nicotinic (nAcChR) and muscarinic (mAcCh) acetylcholine receptors and acetylcholinesterase (AcChEase) are structurally unrelated but share a common functional property: interaction with acetylcholine (AcCh). Alignment of the probable AcCh binding site regions of the nAcChR and mAcChR protein sequences revealed the presence of ten nearly identically spaced consensus residues, six of which contain potentially ligand-interactive side chains. Important elements of the consensus residues also were found in one unique sequence region of the AcChEases. Alignments among the two receptors and AcChEase outside the apparent binding region were rare, and the consensus AcCh binding residues were largely substituted in the homologous proteins, which do not bind AcCh. The consensus residues include two possible anionic subsite Asp residues and a Ser that may hydrogen bond to the AcCh carbonyl in the receptors. These residues correspond to positions Asp-166, Ser-173, and Asp-200 in the neuromuscular nAcChR; Asp-71, Ser-78, and Asp-105 in the M1 mAcChR; and Asp-93 and Asp-128 in Torpedo AcChEase. No corresponding consensus Ser is found in the AcChEase sequence; this is expected because of a downstream esterase active-site Ser-200 (Torpedo). A receptor-conserved and disulfide-linked Cys corresponding to neuromuscular nAcChR residue 193 and M1 mAcChR residue 97 may be important in energy transduction associated with agonist-mediated events. The presence of additional binding-site aromatic residues that may form a hydrophobic environment near the anionic subsite are aligned within, but not between, the three cholinergic protein groups. These observations target specific regions and residues within these proteins for structure-function studies of the cholinergic binding domain.
Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331.
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Peterson
D000109:Acetylcholine / Q000378:metabolism*; D000110:Acetylcholinesterase / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D006801:Humans; D008969:Molecular Sequence Data; D011976:Receptors, Muscarinic / Q000235:genetics* / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D011976:Receptors, Muscarinic; D011978:Receptors, Nicotinic; D000110:Acetylcholinesterase; D000109:Acetylcholine