Abstract
Investigating how a test drug alters the reaction of a site-directed electrophile with a receptor is a powerful method for determining whether the drug acts competitively or allosterically, provided that the binding site of the electrophile is known. In this study, therefore, we mutated nucleophilic residues near and within the orthosteric pockets of M(1) and M(2) muscarinic receptors to identify where acetylcholine mustard and 4-[(2-bromoethyl)methyl-amino]-2-butynyl-N-(3-chlorophenyl)carbamate (BR384) bind covalently. BR384 is the nitrogen mustard analog of [4-[[N-(3-chlorophenyl)carbamoyl]oxy]-2-butynyl]trimethylammonium chloride (McN-A-343). Mutation of the highly conserved aspartic acid in M(1) (Asp105) and M(2) (Asp103) receptors to asparagine largely prevented receptor alkylation by acetylcholine mustard, although modest alkylation still occurred at M(2) D103N at high concentrations of the mustard. Receptor alkylation by BR384 was also greatly inhibited in the M(1) D105N mutant, but some alkylation still occurred at high concentrations of the compound. In contrast, BR384 rapidly alkylated the M(2) D103N mutant. Its affinity was reduced to one tenth, however. The alkylation of M(2) D103N by BR384 was competitively inhibit...Continue Reading
References
Mar 1, 1975·British Journal of Pharmacology·D A RobinsonJ M Young
Jan 1, 1990·Journal of Medicinal Chemistry·B RingdahlD J Jenden
Nov 1, 1972·Journal of Medicinal Chemistry·C H Jackson, M Hirst
Jan 1, 1993·Life Sciences·S LazarenoN J Birdsall
Jan 12, 1999·Archives of Biochemistry and Biophysics·W K VogelM I Schimerlik
Jan 1, 1995·Life Sciences·R D SchwarzA J Thomas
Apr 6, 2000·The Journal of Biological Chemistry·J JakubikS Tucek
Jan 28, 2003·Biochemical Society Transactions·E C HulmeM S Bee
Jun 25, 2005·Mini Reviews in Medicinal Chemistry·N J M Birdsall, S Lazareno
Oct 27, 2007·Science·Vadim CherezovRaymond C Stevens
Aug 7, 2008·The Journal of Pharmacology and Experimental Therapeutics·Hinako SugaFrederick J Ehlert
Aug 30, 2008·The Journal of Biological Chemistry·Celine ValantArthur Christopoulos
Oct 1, 2008·The Journal of Pharmacology and Experimental Therapeutics·Katherine W FigueroaFrederick J Ehlert
Nov 17, 2009·Biochemical Pharmacology·Hinako Suga, Frederick J Ehlert
Jul 2, 2010·British Journal of Pharmacology·K W FigueroaF J Ehlert
Citations
Mar 24, 2016·Cancer Letters·Ying LiuTian-Rui Xu
Mar 3, 2004·Biophysical Journal·Karen A SelzMichael J Owens
Feb 19, 2021·Journal of Medicinal Chemistry·Natalia V Ortiz ZacaríasLaura H Heitman
Jul 13, 2020·Biochemical Pharmacology·Xue YangAdriaan P IJzerman
Oct 24, 2018·Journal of Medicinal Chemistry·Márton VassChris de Graaf
Jul 6, 2013·Biochemistry·Hinako Suga, Frederick J Ehlert