PUBLICATION
Molecular cloning and characterization of an atypical butyrylcholinesterase-like protein in zebrafish
- Authors
- Tan, K.S., Zhang, Y., Liu, L., Li, S., Zou, X., Zeng, W., Cheng, G., Wang, D., Tan, W.
- ID
- ZDB-PUB-210305-10
- Date
- 2021
- Source
- Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology 255: 110590 (Journal)
- Registered Authors
- Keywords
- Butyrylcholinesterase, Molecular docking, Native-PAGE and LC-MS, Neighbor-joining phylogenetic analysis, Zebrafish
- MeSH Terms
-
- Animals
- Carboxylic Ester Hydrolases*/biosynthesis
- Carboxylic Ester Hydrolases*/chemistry
- Carboxylic Ester Hydrolases*/genetics
- Cloning, Molecular*
- Protein Domains
- Zebrafish*/genetics
- Zebrafish*/metabolism
- Zebrafish Proteins*/biosynthesis
- Zebrafish Proteins*/chemistry
- Zebrafish Proteins*/genetics
- PubMed
- 33662568 Full text @ Comp. Biochem. Physiol. B Biochem. Mol. Biol.
Citation
Tan, K.S., Zhang, Y., Liu, L., Li, S., Zou, X., Zeng, W., Cheng, G., Wang, D., Tan, W. (2021) Molecular cloning and characterization of an atypical butyrylcholinesterase-like protein in zebrafish. Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology. 255:110590.
Abstract
Cholinesterases act as bio scavengers to clear organophosphorus (OP) compounds and prodrugs. The butyrylcholinesterase (BChE) gene has been found in several types of teleost fish but this gene has yet to be identified in cyprinid fish. Indeed, BChE homologs have not been found in the zebrafish (Danio rerio) genomic database. Here, we demonstrate that BChE activity is present in zebrafish, in line with other groups' findings. Using in-gel native-PAGE enzymatic activity staining and LC-MS/MS technique, an atypical BChE-like protein was identified in zebrafish. The si:ch211-93f2.1 gene was cloned, and His-tagged recombinant protein was expressed using the Pichia yeast system. The purified protein (molecular weight ~ 180 kDa) showed BChE activity, and degraded acetylcholinesterase (ACh) at a higher rate than BCh. However, phylogram analysis shows that this novel cholinesterase shared an evolutionary origin with carboxylic esterase rather than BChE. The zebrafish BChE-like protein shares structural characteristics with cholinesterase and carboxylesterase. The 2-arachidonoylglycerol (2-AG), nicosulfuron, and triacetin exhibited a higher binding affinity to the zebrafish BChE-like protein than BCh and ACh. With the identification of BChE-like protein in zebrafish, this study could shed light on the origin of BChE and may contribute towards the development of a BChE knockout zebrafish model for sensitive drug or toxin screening.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping