Elucidating the Three-Dimensional Structure of α-dystroglycan using Negative Staining
Naoki Kasahata1,2*
1Institute of Laboratory of Structural Biology, Minamicho 56-1-111, Itabashi-ku, Tokyo, 173-0027, Japan
2Molecular Composite Physiology Research Group, Health and Medical Research Institute, National Institute of Advanced Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8565, Japan
Citation: Kasahata N (2025) Elucidating the Three-Dimensional Structure of ?-dystroglycan using Negative Staining. Appl Cell Biol, 13(1), [14-28]
Abstract
Background: Abnormal glycosylation of a-dystroglycan causes many diseases, such as congenital muscular dystrophies. Determining the a-dystroglycan structure is very important for elucidating the pathogenesis of these disorders and treating them.
Methods: In this study, a-dystroglycan was purified using DEAE (diethylaminoethyl) Sephacel anion-exchange chromatography, WGA (Wheat Germ Agglutinin) affinity chromatography, and size-exclusion chromatography.
Results: The three-dimensional structure of a-dystroglycan was studied using negative staining and single-particle analysis, which revealed two globular domains connected by a flexible linker. The three-dimensional structure of a-dystroglycan is consistent with previously reported 2D images.
Conclusion: Clarifying the three-dimensional structure presented here of a-dystroglycan will help to reveal the molecular mechanisms underlying a-dystroglycanopathy and facilitate its treatment.