title: Analysis of dystrophin in muscular diseases by two-dimensional gel electrophoresis using agarose gels in the first dimension.
abstract: We analyzed dystrophin in case of normal control, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) and infectious muscular disease using two-dimensional gel electrophoresis and immunoblotting with 3 monoclonal dystrophin antibodies: Dys 1, a mid-rod-domain antibody; Dys 2, a C-terminal-domain antibody; and Dys 3, an N-terminal-domain antibody. In cases of normal control, a clearly separated doublet of bands was observed for Dys 1 and 3 at molecular weights 400 and 420 kDa. The isoelectric point was between pH approximately 5.7-approximately 5.9, similar to that for the myosin heavy chain. In one DMD case, a single faint band was observed for Dys 2. BMD presented a single-band pattern for each antibody. Infectious diseases cases showed 3- to 5-band patterns for Dys 1 and single or no bands for Dys 2 and 3. The pI of the Dys 1 band was almost identical. These results suggest coexistence of normal dystrophin and its proteolytic products, both containing triple helical segment, and show that two-dimensional gel electrophoresis may be applicable in the analysis of dystrophin in muscular disease.
authors.affiliation: Department of Pediatrics, University of Tsukuba, Ibaraki-ken, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Hamano
authors.forename: T
authors.initials: T
authors.lastname: Takeya
authors.forename: N
authors.initials: N
authors.lastname: Iwasaki
authors.forename: N
authors.initials: N
authors.lastname: Okoshi
authors.forename: T
authors.initials: T
authors.lastname: Fukubayashi
authors.forename: M
authors.initials: M
authors.lastname: Kirinoki
authors.forename: Y
authors.initials: Y
authors.lastname: Yao
authors.forename: T
authors.initials: T
authors.lastname: Hirabayashi
authors.forename: H
authors.initials: H
authors.lastname: Takita
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D003882:Dermatomyositis / Q000378:metabolism; D016189:Dystrophin / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D007223:Infant; D008875:Middle Aged; D009132:Muscles / Q000737:chemistry*; D009136:Muscular Dystrophies / Q000378:metabolism*; D009220:Myositis / Q000378:metabolism*
chemical_list: D016189:Dystrophin