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A direct method to visualise the aryl acylamidase activity on cholinesterases in polyacrylamide gels
BACKGROUND: In vertebrates, two types of cholinesterases exist, acetylcholinesterase and butyrylcholinesterase. The function of acetylcholinesterase is to hydrolyse acetylcholine, thereby terminating the neurotransmission at cholinergic synapse, while the precise physiological function of butyrylcho...
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2000
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC29059/ https://www.ncbi.nlm.nih.gov/pubmed/11231883 http://dx.doi.org/10.1186/1471-2091-1-3 |
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author | Jaganathan, Lakshmanan Boopathy, Rathanam |
author_facet | Jaganathan, Lakshmanan Boopathy, Rathanam |
author_sort | Jaganathan, Lakshmanan |
collection | PubMed |
description | BACKGROUND: In vertebrates, two types of cholinesterases exist, acetylcholinesterase and butyrylcholinesterase. The function of acetylcholinesterase is to hydrolyse acetylcholine, thereby terminating the neurotransmission at cholinergic synapse, while the precise physiological function of butyrylcholinesterase has not been identified. The presence of cholinesterases in tissues that are not cholinergically innervated indicate that cholinesterases may have functions unrelated to neurotransmission. Furthermore, cholinesterases display a genuine aryl acylamidase activity apart from their predominant acylcholine hydrolase activity. The physiological significance of this aryl acylamidase activity is also not known. The study on the aryl acylamidase has been, in part hampered by the lack of a specific method to visualise this activity. We have developed a method to visualise the aryl acylamidase activity on cholinesterase in polyacrylamide gels. RESULTS: The o-nitroaniline liberated from o-nitroacetanilide by the action of aryl acylamidase activity on cholinesterases, in the presence of nitrous acid formed a diazonium compound. This compound gave an azo dye complex with N-(1-napthyl)-ethylenediamine, which appeared as purple bands in polyacrylamide gels. Treating the stained gels with trichloroacetic acid followed by Tris-HCl buffer helped in fixation of the stain in the gels. By using specific inhibitors for acetylcholinesterase and butyrylcholinesterase, respectively, differential staining for the aryl acylamidase activities on butyrylcholinesterase and acetylcholinesterase in a sample containing both these enzymes has been demonstrated. A linear relationship between the intensity of colour developed and activity of the enzyme was obtained. CONCLUSIONS: A novel method to visualise the aryl acylamidase activity on cholinesterases in polyacrylamide gels has been developed. |
format | Text |
id | pubmed-29059 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2000 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-290592001-03-22 A direct method to visualise the aryl acylamidase activity on cholinesterases in polyacrylamide gels Jaganathan, Lakshmanan Boopathy, Rathanam BMC Biochem Methodology Article BACKGROUND: In vertebrates, two types of cholinesterases exist, acetylcholinesterase and butyrylcholinesterase. The function of acetylcholinesterase is to hydrolyse acetylcholine, thereby terminating the neurotransmission at cholinergic synapse, while the precise physiological function of butyrylcholinesterase has not been identified. The presence of cholinesterases in tissues that are not cholinergically innervated indicate that cholinesterases may have functions unrelated to neurotransmission. Furthermore, cholinesterases display a genuine aryl acylamidase activity apart from their predominant acylcholine hydrolase activity. The physiological significance of this aryl acylamidase activity is also not known. The study on the aryl acylamidase has been, in part hampered by the lack of a specific method to visualise this activity. We have developed a method to visualise the aryl acylamidase activity on cholinesterase in polyacrylamide gels. RESULTS: The o-nitroaniline liberated from o-nitroacetanilide by the action of aryl acylamidase activity on cholinesterases, in the presence of nitrous acid formed a diazonium compound. This compound gave an azo dye complex with N-(1-napthyl)-ethylenediamine, which appeared as purple bands in polyacrylamide gels. Treating the stained gels with trichloroacetic acid followed by Tris-HCl buffer helped in fixation of the stain in the gels. By using specific inhibitors for acetylcholinesterase and butyrylcholinesterase, respectively, differential staining for the aryl acylamidase activities on butyrylcholinesterase and acetylcholinesterase in a sample containing both these enzymes has been demonstrated. A linear relationship between the intensity of colour developed and activity of the enzyme was obtained. CONCLUSIONS: A novel method to visualise the aryl acylamidase activity on cholinesterases in polyacrylamide gels has been developed. BioMed Central 2000-12-20 /pmc/articles/PMC29059/ /pubmed/11231883 http://dx.doi.org/10.1186/1471-2091-1-3 Text en Copyright © 2000 Jaganathan and Boopathy; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL. |
spellingShingle | Methodology Article Jaganathan, Lakshmanan Boopathy, Rathanam A direct method to visualise the aryl acylamidase activity on cholinesterases in polyacrylamide gels |
title | A direct method to visualise the aryl acylamidase activity on cholinesterases in polyacrylamide gels |
title_full | A direct method to visualise the aryl acylamidase activity on cholinesterases in polyacrylamide gels |
title_fullStr | A direct method to visualise the aryl acylamidase activity on cholinesterases in polyacrylamide gels |
title_full_unstemmed | A direct method to visualise the aryl acylamidase activity on cholinesterases in polyacrylamide gels |
title_short | A direct method to visualise the aryl acylamidase activity on cholinesterases in polyacrylamide gels |
title_sort | direct method to visualise the aryl acylamidase activity on cholinesterases in polyacrylamide gels |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC29059/ https://www.ncbi.nlm.nih.gov/pubmed/11231883 http://dx.doi.org/10.1186/1471-2091-1-3 |
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