| Autor: |
Wang Y; Department of Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, China., Zhang JY; Department of Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, China., Teng JY; Department of Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, China., Xiong HF; Department of Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, China., Li QF; Department of Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, China. |
| Jazyk: |
angličtina |
| Zdroj: |
The Journal of veterinary medical science [J Vet Med Sci] 2023 Feb 21; Vol. 85 (2), pp. 244-251. Date of Electronic Publication: 2023 Jan 02. |
| DOI: |
10.1292/jvms.22-0222 |
| Abstrakt: |
Locoweeds, a type of poisonous weedare, are widely distributed throughout the world and have a significant impact on the development of herbivore animal husbandry. Swainsonine (SW), the main toxin in locoweeds, can competitively inhibit lysosomes α-mannosidase (LAM) in animal cells, resulting in α-mannosidosis. However, the specifics of the interaction between SW and LAM are still unclear. Here, we used molecular docking to predicte the interaction points between SW and LAM, built mutated lysosomes α-mannosidase (LAM M ), and analyzed its biochemical properties changes in presumption points. The Trp at the 28th position and the Tyr at the 599th position of the LAM were interaction point candidates, and the above two amino acids in Capra hircus LAM (chLAM), were successfully mutated to glycine by constructing recombinant yeast GS115/PIC9K- LAM M . The results showed that the sensitivity of Capra hircus LAM M (chLAM M ), to SW decreased significantly compared with wild-type LAM, the enzyme activity of LAM decreased approximately threefold, the optimum temperature of LAM M decreased from 55°C to 50°C, the optimum pH value increased from 4.5 to 5.0, and the effects of Mn 2+ , Fe 3+ , Al 3+ , Co 2+ , Cr 3+ , and ethylenediaminetetraacetic acid (EDTA) on LAM enzyme activity before and after point mutation changed significantly. These findings help us better understanding the molecular mechanism of the interaction mechanism between SW and chLAM, and provide new reference for solving locoweeds poisoning. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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