The design of a new truncated and engineered alpha1-antitrypsin based on theoretical studies: an antiprotease therapeutics for pulmonary diseases.

ABSTRACT: Alpha 1- antitrypsin (?1AT) a 54 kDa glycoprotein is a protease inhibitor. In the absence of ?1AT, elastase released by lung macrophages, was not inhibited and lead to elastin breakdown and pulmonary problems such as emphysema or COPD. ?1AT has three site of N-glycosylation and a characteristic reactive central loop (RCL). As small-scale medicines are preferred for pulmonary drug delivery, in this study ?1ATs (1, 2, 3, 4 and 5) were engineered and shortened from the N-terminal region. In order to investigate the effect of different mutations and the deletion of 46 amino acids theoretical studies were performed. Homology modeling was performed to generate the 3D structure of ?1ATs. The 10 ns Molecular Dynamic (MD) simulations were carried out to refine the models. Results from MD and protein docking showed that ?1AT2 has the highest binding affinity for neutrophil elastase, provided the basis for the experimental phase in which sequences from the five ?1AT constructs were inserted into the expression vector pGAPZ? and expressed in the yeast Pichia pastoris. Although, the ?1AT2 construct has the highest inhibitory activity even more that the native construct (?1AT5), results indicated the presence of protease inhibitory function of all the proteins' construct against elastase.

SUBMITTER: Pirooznia N 

PROVIDER: S-EPMC3698207 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature