Journal
HUMAN MOLECULAR GENETICS
Volume 20, Issue 15, Pages 2955-2963Publisher
OXFORD UNIV PRESS
DOI: 10.1093/hmg/ddr199
Keywords
-
Funding
- NIH [AR007612]
- National Institute of Arthritis, Musculoskeletal and Skin Diseases [AR042423]
- Muscular Dystrophy Association
- Nash Avery Foundation
- Charley's Fund
Ask authors/readers for more resources
Duchenne muscular dystrophy (DMD) is a deadly and common childhood disease caused by mutations that disrupt dystrophin protein expression. Several miniaturized dystrophin/utrophin constructs are utilized for gene therapy, and while these constructs have shown promise in mouse models, the functional integrity of these proteins is not well described. Here, we compare the biophysical properties of full-length dystrophin and utrophin with therapeutically relevant miniaturized constructs using an insect cell expression system. Full-length utrophin, like dystrophin, displayed a highly cooperative melting transition well above 37 degrees C. Utrophin constructs involving N-terminal, C-terminal or internal deletions were remarkably stable, showing cooperative melting transitions identical to full-length utrophin. In contrast, large dystrophin deletions from either the N- or C-terminus exhibited variable stability, as evidenced by melting transitions that differed by 20 degrees C. Most importantly, deletions in the large central rod domain of dystrophin resulted in a loss of cooperative unfolding with increased propensity for aggregation. Our results suggest that the functionality of dystrophin therapeutics based on mini-or micro-constructs may be compromised by the presence of non-native protein junctions that result in protein misfolding, instability and aggregation.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available