| Full text | |
| Author(s): |
Lopes Vilela, Adriana Ferreira
[1]
;
Seidl, Claudia
[1]
;
Lima, Juliana Maria
[1]
;
Cardoso, Carmen Lucia
[1]
Total Authors: 4
|
| Affiliation: | [1] Univ Sao Paulo, Dept Quim, Grp Cromatog Bioafinidade & Prod Nat, Fac Filosofia Ciencias & Letras Ribeirao Preto, BR-14040901 Ribeirao Preto, SP - Brazil
Total Affiliations: 1
|
| Document type: | Journal article |
| Source: | Analytical Biochemistry; v. 549, p. 53-57, MAY 15 2018. |
| Web of Science Citations: | 7 |
| Abstract | |
Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are key cholinesterase enzymes responsible for the hydrolysis of acetylcholine into choline and acetic acid, an essential process for the restoration of the cholinergic neuron. The loss of cholinergic function in the central nervous system contributes to the cognitive decline associated with advanced age and Alzheimer s disease (AD). Inhibitions assays represent a significant role in the drug discovery process Herein, we describe an improved label free method to screen and characterize new BChE ligands. The liquid chromatography system uses an immobilized capillary enzyme reactor (ICER) as a low affinity and high selectivity column coupled to a mass spectrometer (MS). The enzyme activity was evaluated by monitoring the choline's precursor ion {[}M + H](+) m/z 104 for a brief period. The method was validated using two known cholinesterase inhibitors tacrine and galanthamine. The IC50 values were 0 03 +/- 0.006 mu M and 0.88 +/- 0.2 for tacrine and galanthamine respectively, and Ki was 0.11 +/- 0.2 for galanthamine. The efficient combination of the ftuBChE-ICER with sensitive enzymatic assay detection such as MS, improved the reliable, fast identification of new ligands. Moreover, specific direct quantitation of the product contributes to the reduction of false positive and negative results. (AU) | |
| FAPESP's process: | 16/02873-0 - Acetylcholinesterase and beta-secretase 1: a study of co-immobilization conditions for screening of ligands |
| Grantee: | Adriana Ferreira Lopes Vilela |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 14/11640-3 - LC-MS / 2D methods for screening ligands for evaluating enzyme inhibitory activity and identification of active compounds in crude extracts |
| Grantee: | Cláudia Seidl |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |
| FAPESP's process: | 14/06907-0 - Immobilization nucleoside diphosphate kinase (NDK) enzyme Leishmania major: the study of conditions for the application in the screening of ligands |
| Grantee: | Juliana Maria de Lima |
| Support Opportunities: | Scholarships in Brazil - Doctorate (Direct) |