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GOLGI FRACTIONS PREPARED FROM RAT LIVER HOMOGENATES
Open accessAbstract
In devising a new procedure for the isolation of Golgi fractions from rat liver homogenates, we have taken advantage of the overloading with very low density lipoprotein (VLDL) particles that occurs in the Golgi elements of hepatocytes ∼90 min after ethanol is administered (0.6 g/100 g body weight) by stomach tube to the animals. The VLDLs act as morphological markers as well as density modifiers of these elements. The starting preparation is a total microsomal fraction prepared from liver homogenized (1:5) in 0.25 M sucrose. This fraction is resuspended in 1.15 M sucrose and loaded at the bottom of a discontinuous sucrose density gradient. Centrifugation at ∼13 x 106 g·min yields by flotation three Golgi fractions of density >1.041 and <1.173. The light and intermediate fractions consist essentially of VLDL-loaded Golgi vacuoles and cisternae. Nearly empty, often collapsed, Golgi cisternae are the main component of the heavy fraction. A procedure which subjects the Golgi fractions to hypotonic shock and shearing in a French press at pH 8.5 allows the extraction of the content of the Golgi elements and the subsequent isolation of their membranes by differential centrifugation.
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Cited by
Roth, J., & Berger, E. G. (1982) · The Journal of cell biology · doi:10.1083/jcb.93.1.223 · Cited by 619
Borchardt, R., & Davis, R. (1987) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)49269-1 · Cited by 304
The mannose-6-phosphate receptor for lysosomal enzymes is concentrated in cis Golgi cisternae
Brown, W. J., & Farquhar, M. G. (1984) · Cell · doi:10.1016/0092-8674(84)90223-x · Cited by 288
Apolipoprotein E localization in rat hepatocytes by immunogold labeling of cryothin sections
Hamilton, R., Wong, J., Guo, L., Krisans, S., & Havel, R. (1990) · Journal of Lipid Research · doi:10.1016/s0022-2275(20)42343-0 · Cited by 129
Cryo-EM and artificial intelligence visualize endogenous protein community members
Skalidis, I., Kyrilis, F. L., Tüting, C., Hamdi, F., Chojnowski, G., & Kastritis, P. L. (2022) · Structure · doi:10.1016/j.str.2022.01.001 · Cited by 61
Metadata
- Authors
- Ehrenreich, J. H., Bergeron, J. J. M., Siekevitz, P., & Palade, G. E.
- Venue
- The Journal of Cell Biology
- Publisher
- Rockefeller University Press
- Published
- 1973-10-01
- Volume
- 59
- Issue
- 1
- Pages
- 45-72
- ISSN
- 0021-9525 (ISSN-L)
- 1540-8140
- OpenAlex
- W1997321425
- Cited by
- 448
- References
- 0
- FWCI
- 15.14
- Type
- journal article
- Language
- en
- Open access
- green
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- Ehrenreich, J. H., Bergeron, J. J. M., Siekevitz, P., & Palade, G. E. (1973). GOLGI FRACTIONS PREPARED FROM RAT LIVER HOMOGENATES. The Journal of Cell Biology. https://doi.org/10.1083/jcb.59.1.45
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
