In vitro expressed dystrophin fragments do not associate with each other

FEBS Letters

Kunkel, Louis M.¹  Chan, Yiu-mo¹ 

[1] Howard Hughes Medical Institute, Enders 570, and Division of Genetics at Children's Hospital, 300 Longwood Avenue, Boston, MA 02115, USA

关键词: Dystrophin;    In vitro translation;    Dimerization;    Immunoprecipitation;    Blot overlay;   

DOI  :  10.1016/S0014-5793(97)00454-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Dystrophin, a component of the muscle membrane cytoskeleton, is the protein altered in Duchenne Muscular Dystrophy (DMD) and Becker Muscular Dystrophy (BMD). Dystrophin shares significant homology with other cytoskeletal proteins, such as α-actinin and spectrin. On the basis of its sequence similarity with α-actinin and spectrin, dystrophin has been proposed to function as dimer. However, the existence of both dimers and monomers have been observed by electron microscopy. To address this apparent discrepancy, we expressed dystrophin fragments composed of different domains in an in vitro translation system. The expressed fragments were tested for their ability to interact with each other and full-length dystrophin by both immunoprecipitation and blot overlay assays. These assays were successfully used to demonstrate the dimerization of α-actinin and spectrin, yet failed to detect any interaction between dystrophin fragments. Although these in vitro results do not prove that dystrophin is not a dimer in vivo, they do indicate that this interaction is not like that of the α-actinin and spectrin.

【 授权许可】

Unknown