RESUMEN

Lysosomal α-Mannosidase (LAM) belongs to the glycoside hydrolyzing enzymes family 38 and is involved in the biosynthesis and turnover of N-linked glycoproteins process. Locoweeds, which contain swainsonine (SW) that inhibits LAM, are the main poisoning plants in many regions of the world, and thereby resulting in animal poisoning or even death. Based on regions of protein sequence conservation between LAM from Bos taurus and Homo sapiens, we cloned cDNA encoding Capra hircus LAM (chLAM). Expression of cDNA in Pichia pastoris resulted in the secretion of aLAM activity into the culture medium. The recombinant chLAM was activated 1.6 and 1.2-fold with Zn(2+) and Ca(2+), respectively. By homology modeling, molecular docking and mutant analysis, we obtained the probable binding modes of SW at the allosteric sites of chLAM, and the potential mutant sites for the resistance to SW. Prediction of SW sensitivity to A28 W/G, D58 Y/G mutant chLAM is lower than wild type chLAM. The obtained results lead to a better understanding of not only interactions between substrate/SW and chLAM, but also of a potential strategy for a novel therapy for locoweed poisoning.

Asunto(s)

Glicoproteínas/genética , Lisosomas/enzimología , Oxytropis/envenenamiento , alfa-Manosidasa/genética , Secuencia de Aminoácidos , Animales , Bovinos , Clonación Molecular , Secuencia Conservada/genética , Cabras/genética , Humanos , Mutación , Oxytropis/genética , Pichia , alfa-Manosidasa/química

RESUMEN

@#Objective To study the clinical efficacy of electromyography biofeedback on dystonia after hepatolenticular degeneration.Methods 40 patients with dystonia after hepatolenticular degeneration were divided into treatment group (n=20) and control group (n=20).All patients were treated with copper-cleaning, clonazepam and benzhexol hydrochloride etc., and acupuncture. The treatment group wastreated with electromyographic biofeedback in addition. They were assessed with modified Ashworth scale, modified Barthel index, and ankledorsiflexion active range of motion. Results The lower extremities function of all patients improved after treatment (P<0.05) and thetreatment group improved more than the control group (P<0.05). Conclusion Electromyography biofeedback is more effective to improvethe lower extremities function and activity of daily living in patients with hepatolenticular degeneration following dystonia.