Study of glial fibrillary acidic protein in a human glioma cell line grown in culture and as a solid tumor.
Journal Article
A continuous human glioma cell line grown in culture and as a solid tumor was analyzed for glial fibrillary acidic (GFA) protein. This material provided a rich source for GFA protein that could also be manipulated and controlled. Immunoperoxidase staining at the light and electron microscopic levels revealed that the cell culture and tumor specimens were strongly positive for GFA protein. When aqueous soluble fractions of the cell culture and tumor were separated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, electroblotted onto nitrocellulose, and stained immunochemically, they contained exclusively low molecular weight (41--43K-dalton) GFA peptides. SDS (0.15%)-soluble fractions contained either low molecular weight only (culture) or a mixture of peptides ranging from 41 to 49K daltons. SDS (1%) extracts of either cell culture or tumor contained only 49K-dalton GFA protein. Two-dimensional gel separation revealed that the GFA protein extracted from either the culture or tumor with 1% SDS resolved to two or three spots at pH 5.8. Low molecular weight GFA peptides (less than 49K daltons) in aqueous and 0.15% SDS-soluble extracts became increasingly more acidic with decreasing molecular weight. The extremely rapid degradation seen suggests that this cell line may be a valuable system for further study of intermediate filament protein turnover.
Full Text
Duke Authors
Cited Authors
- Bigbee, JW; Bigner, DD; Pegram, C; Eng, LF
Published Date
- February 1, 1983
Published In
Volume / Issue
- 40 / 2
Start / End Page
- 460 - 467
PubMed ID
- 6822832
Pubmed Central ID
- 6822832
International Standard Serial Number (ISSN)
- 0022-3042
Digital Object Identifier (DOI)
- 10.1111/j.1471-4159.1983.tb11305.x
Language
- eng
Conference Location
- England