Insertion of a myc-tag within α-dystroglycan domains improves its biochemical and microscopic detection.

ABSTRACT:

Background

Epitope tags and fluorescent fusion proteins have become indispensable molecular tools for studies in the fields of biochemistry and cell biology. The knowledge collected on the subdomain organization of the two subunits of the adhesion complex dystroglycan (DG) enabled us to insert the 10 amino acids myc-tag at different locations along the α-subunit, in order to better visualize and investigate the DG complex in eukaryotic cells.

Results

We have generated two forms of DG polypeptides via the insertion of the myc-tag 1) within a flexible loop (between a.a. 170 and 171) that separates two autonomous subdomains, and 2) within the C-terminal domain in position 500. Their analysis showed that double-tagging (the β-subunit is linked to GFP) does not significantly inte

SUBMITTER: Morlacchi S 

PROVIDER: S-EPMC3432625 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature