Expression and subcellular localization of P9-ZFD protein in patients with myasthenia gravis.
Chin Med Sci J
; 19(3): 221-4, 2004 Sep.
Article
in En
| MEDLINE
| ID: mdl-15506653
ABSTRACT
OBJECTIVE:
To express and purify the protein coded by the TRAF-type zinc finger domain of myasthenia gravis (MG)-related gene P9 (P9-ZFD) and to prepare P9-ZFD antiserum for detecting expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of patient with MG.METHODS:
The cDNA encoding P9-ZFD was amplified by RT-PCR. The cloned P9-ZFD cDNA was ligated into pET24a, and the P9-ZFD recombinant protein was induced via E. coli. BL21 (DE3) and purified by histidine affinity chromatography. P9-ZFD antiserum was prepared and its titer and specificity were determined by ELISA and Western blot. Expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of MG and control were studied.RESULTS:
The molecular weight of purified P9-ZFD protein was about 30 kD. Its purity was more than 95%. Antiserum specific for P9-ZFD was excellent. P9-ZFD protein is fully confined to the cytoplasm membrane of skeletal muscle cell of MG, obvious immunostaining was absent in the A, I, and Z bands of cytoplasm and no immunoreactivity was observed in the skeletal muscle cell of control.CONCLUSION:
P9-ZFD protein is expressed as a cytoplasm membrane-bound protein and has obvious distribution difference in the skeletal muscle cells of patient with MG and normal control.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Muscle, Skeletal
/
Muscle Proteins
/
Myasthenia Gravis
Limits:
Adult
/
Female
/
Humans
Language:
En
Journal:
Chin Med Sci J
Journal subject:
TERAPIAS COMPLEMENTARES
Year:
2004
Document type:
Article