1-Naphthyl acetate as an alternative substrate of hemolysate cholinesterase: Direct visualization of enzyme activity within 10 minutes on polyacrylamide gels
IR@NISCAIR: CSIR-NISCAIR, New Delhi - ONLINE PERIODICALS REPOSITORY (NOPR)
View Archive Info| Field | Value | |
| Title |
1-Naphthyl acetate as an alternative substrate of hemolysate cholinesterase: Direct visualization of enzyme activity within 10 minutes on polyacrylamide gels
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| Creator |
Chowdhary, Sheemona
Bhattacharyya, Rajasri Banerjee, Dibyajyoti |
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| Subject |
Acetylcholinesterase
Biomarker Karnovsky staining Naphthyl acetates Organophosphorus pesticides |
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| Description |
662-671
Hemolysate cholinesterase is currently recognized as the most preferred biomarker to detect acute organophosphorus poisoning. Direct visualization of cholinesterase activity on polyacrylamide gels is routinely practiced using acetylthiocholine as a substrate. Overnight incubation with the staining solution is required to understand the enzyme activity bands on gels. Therefore, the need arises to explore rapid detection methods, which can specifically detect hemolysate cholinesterase on polyacrylamide gels. Here, we have explored alternative substrates, such as 1-NA and 2-NA which might have the potential to behave as specific substrates for the detection of hemolysate cholinesterase activity on the gels. It is observed by the <em>in silico</em> studies that 1-NA bind at the active site of acetylcholinesterase akin to acetylcholine (ACh) with a better fitness score. Secondly, the hemolysate cholinesterase activity, as well as its inhibition by organophosphorus pesticides is understandable within 10 min using Fast Blue RR dye for the detection of 1-NA. The organophosphorus inhibited activity is regained in the presence of cholinesterase reactivator. Moreover, the enzyme activity bands formed using 1-NA proves the specificity of the substrate for hemolysate cholinesterase as in the presence of specific acetylcholinesterase inhibitors the band formation disappears. On the other hand, ATCh requires minimum 8-12 h staining time for detection of enzyme activity band following Karnovsky and Roots protocol. Our results prove that 1-NA is an alternative substrate of hemolysate cholinesterase which specifically detects the enzyme activity on gel rapidly. We recommend 1-NA for rapid detection of hemolysate cholinesterase activity on the gels. |
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| Date |
2019-09-04T06:29:57Z
2019-09-04T06:29:57Z 2019-09 |
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| Type |
Article
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| Identifier |
0975-1009 (Online); 0019-5189 (Print)
http://nopr.niscair.res.in/handle/123456789/50453 |
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| Language |
en_US
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| Rights |
<img src='http://nopr.niscair.res.in/image/cc-license-sml.png'> <a href='http://creativecommons.org/licenses/by-nc-nd/2.5/in' target='_blank'>CC Attribution-Noncommercial-No Derivative Works 2.5 India</a>
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| Publisher |
NISCAIR-CSIR, India
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| Source |
IJEB Vol.57(09) [September 2019]
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